Mesenchymal Stem Cells Transplantation Group Research Articles

Efficacy of Neonatal Porcine Bone Marrow-Derived Mesenchymal Stem Cell Xenotransplantation for the Therapy of Hind Limb Lymphedema in Mice.

Lymphedema is an intractable disease with few effective therapeutic options. Autologous mesenchymal stem cell (MSC) transplantation is a promising therapy for this disease. However, its use is limited by the cost and time for preparation. Recently, xenotransplantation of porcine MSCs has emerged as an alternative to autologous MSC transplantation. In this study, we aimed to clarify the usefulness of neonatal porcine bone marrow-derived MSC (NpBM-MSC) xenotransplantation for the treatment of lymphedema. One million NpBM-MSCs were xenotransplanted into the hind limbs of mice with severe lymphedema (MSC transplantation group). The therapeutic effects were assessed by measuring the femoral circumference, the volume of the hind limb, the number and diameter of lymphatic vessels in the hind limb, and lymphatic flow using a near-infrared fluorescence (NIRF) imaging system. We compared the effects using mice with lymphedema that did not undergo NpBM-MSC transplantation (negative control group). The condition of the transplanted NpBM-MSCs was also evaluated histologically. The femoral circumference and volume of the hind limb had been normalized by postoperative day (POD) 14 in the MSC transplantation group, but not in the negative control group (P = 0.041). NIRF imaging revealed that lymphatic flow had recovered in the MSC transplantation group by POD 14, as shown by an increase in luminance in the hind limb. Histological assessment also showed that the xenotransplantation of NpBM-MSC increased the proliferation of lymphatic vessels, but they had been rejected by POD 14. The xenotransplantation of NpBM-MSCs is an effective treatment for lymphedema, and this is mediated through the promotion of lymphangiogenesis.

Local transplantation of mesenchymal stem cells improves encephalo-myo-synangiosis-mediated collateral neovascularization in chronic brain ischemia

BackgroundTo explore whether local transplantation of mesenchymal stem cells (MSCs) in temporal muscle can promote collateral angiogenesis and to analyze its main mechanisms of promoting angiogenesis.MethodsBilateral carotid artery stenosis (BCAS) treated mice were administrated with encephalo-myo-synangiosis (EMS), and bone marrow mesenchymal stem cells (BMSCs) were transplanted into the temporal muscle near the cerebral cortex. On the 30th day after EMS, the Morris water maze, immunofluorescence, laser speckle imaging, and light sheet microscopy were performed to evaluate angiogenesis; In addition, rats with bilateral common carotid artery occlusion were also followed by EMS surgery, and BMSCs from GFP reporter rats were transplanted into the temporal muscle to observe the survival time of BMSCs. Then, the concentrated BMSC-derived conditioned medium (BMSC-CM) was used to stimulate HUVECs and BMECs for ki-67 immunocytochemistry, CCK-8, transwell and chick chorioallantoic membrane assays. Finally, the cortical tissue near the temporal muscle was extracted after EMS, and proteome profiler (angiogenesis array) as well as RT-qPCR of mRNA or miRNA was performed.ResultsThe results of the Morris water maze 30 days after BMSC transplantation in BCAS mice during the EMS operation, showed that the cognitive impairment in the BCAS + EMS + BMSC group was alleviated (P < 0.05). The results of immunofluorescence, laser speckle imaging, and light sheet microscopy showed that the number of blood vessels, blood flow and astrocytes increased in the BCAS + EMS + BMSC group (P < 0.05). The BMSCs of GFP reporter rats were applied to EMS and showed that the transplanted BMSCs could survive for up to 14 days. Then, the results of ki-67 immunocytochemistry, CCK-8 and transwell assays showed that the concentrated BMSC-CM could promote the proliferation and migration of HUVECs and BMECs (P < 0.05). Finally, the results of proteome profiler (angiogenesis array) in the cerebral cortex showed that the several pro-angiogenesis factors (such as MMP-3, MMP-9, IGFBP-2 or IGFBP-3) were notably highly expressed in MSC transplantation group compared to others.ConclusionsLocal MSCs transplantation together with EMS surgery can promote angiogenesis and cognitive behavior in chronic brain ischemia mice. Our study illustrated that MSC local transplantation can be the potential therapeutical option for improving EMS treatment efficiency which might be translated into clinical application.

Mesenchymal stem cells transplantation combined with IronQ attenuates ICH-induced inflammation response via Mincle/syk signaling pathway

BackgroundIntracerebral hemorrhage (ICH) is a severe brain-injured disease accompanied by cerebral edema, inflammation, and subsequent neurological deficits. Mesenchymal stem cells (MSCs) transplantation has been used as a neuroprotective therapy in nervous system diseases because of its anti-inflammatory effect. Nevertheless, the biological characteristics of transplanted MSCs, including the survival rate, viability, and effectiveness, are restricted because of the severe inflammatory response after ICH. Therefore, improving the survival and viability of MSCs will provide a hopeful therapeutic efficacy for ICH. Notably, the biomedical applications of coordination chemistry-mediated metal-quercetin complex have been verified positively and studied extensively, including growth-promoting and imaging probes. Previous studies have shown that the iron-quercetin complex (IronQ) possesses extraordinary dual capabilities with a stimulating agent for cell growth and an imaging probe by magnetic resonance imaging (MRI). Therefore, we hypothesized that IronQ could improve the survival and viability of MSCs, displaying the anti-inflammation function in the treatment of ICH while also labeling MSCs for their tracking by MRI. This study aimed to explore the effects of MSCs with IronQ in regulating inflammation and further clarify their potential mechanisms.MethodsC57BL/6 male mice were utilized in this research. A collagenase I-induced ICH mice model was established and randomly separated into the model group (Model), quercetin gavage group (Quercetin), MSCs transplantation group (MSCs), and MSCs transplantation combined with IronQ group (MSCs + IronQ) after 24 h. Then, the neurological deficits score, brain water content (BWC), and protein expression, such as TNF-α, IL-6, NeuN, MBP, as well as GFAP, were investigated. We further measured the protein expression of Mincle and its downstream targets. Furthermore, the lipopolysaccharide (LPS)-induced BV2 cells were utilized to investigate the neuroprotection of conditioned medium of MSCs co-cultured with IronQ in vitro.ResultsWe found that the combined treatment of MSCs with IronQ improved the inflammation-induced neurological deficits and BWC in vivo by inhibiting the Mincle/syk signaling pathway. Conditioned medium derived from MSCs co-cultured with IronQ decreased inflammation, Mincle, and its downstream targets in the LPS-induced BV2 cell line.ConclusionsThese data suggested that the combined treatment exerts a collaborative effect in alleviating ICH-induced inflammatory response through the downregulation of the Mincle/syk signaling pathway following ICH, further improving the neurologic deficits and brain edema.

Effects of umbilical cord mesenchymal stem cells on expression of CYR61, FSH, and AMH in mice with premature ovarian failure.

This study aimed to investigate the effects of umbilical cord mesenchymal stem cells on the expression of CYR61, FSH and AMH in mice with premature ovarian failure. For this purpose, thirty SPF female SD mice were selected as the research object, 10 of which were control group, namely group α, and 20 mice with premature ovarian failure model were established by cyclophosphamide. The mice were divided into the model group, namely the β group and the umbilical cord mesenchymal stem cell transplantation group (γ group), with 10 mice in each group. ELSA method was used to determine the levels of follicle-stimulating hormone (FSH), Luteinizing hormone (LH), estradiol (Estradiol) in serum. The changes of E2, Antimullerian hormone (AMH) and cysteine-rich protein 61 in ovarian tissues were determined by the protein imprinting method. Connective tissue growth factor (CTGF) and caspase-3 protein expression. Results showed that in fertility rate, γ group > α group > β group, the difference was statistically significant (P<0.05), in litter size, α group > γ group > β group, the difference was statistically significant (P<0.05). The levels of serum E2 and AMH in α group > γ group > β group, and the levels of serum FSH and LH in β group > γ group > α group were statistically significant (P<0.05). The growth follicles were α group > γ group > β group, and the atresia follicles were β group > γ group > α group, and there was a statistical difference among all groups (P<0.05). There was no difference in luteal number among the three groups (P>0.05). In terms of CYR61 and CTGF protein expression, α group > γ group > β group, and in terms of caspase-3, β group > γ group > α group had statistical significance (P<0.05). It is concluded that intervention with umbilical cord mesenchymal stem cells can significantly improve the expression levels of CYR61 and AMH, reduce the level of FSH, promote cell survival, improve the reproductive quality of mice, and restore the physiological function of the ovary. It is feasible to treat premature ovarian failure with umbilical cord mesenchymal stem cells.

Effects of umbilical cord mesenchymal stem cells on expression of CYR61, FSH and AMH in mice with premature ovarian failure.

The aim of this study was to objective to investigate the effects of umbilical cord mesenchymal stem cells on the expression of CYR61, FSH and AMH in mice with premature ovarian failure. For this purpose, thirty SPF female SD mice were selected as the research object, 10 of which were control group, namely group α, and 20 mice with premature ovarian failure model were established by cyclophosphamide. The mice were divided into model group, namely β group and the umbilical cord mesenchymal stem cell transplantation group (γ group), with 10 mice in each group. ELSA method was used to determine the levels of follicle-stimulating hormone (FSH), Luteinizing hormone (LH), estradiol (Estradiol) in serum. The changes of E2, Antimullerian hormone (AMH) and cysteine-rich protein 61 in ovarian tissues were determined by the protein imprinting method. Connective tissue growth factor (CTGF) and caspase-3 protein expression. Results showed that in fertility rate, γ group > α group > β group, the difference was statistically significant (P<0.05), in litter size, α group > γ group > β group, the difference was statistically significant (P<0.05). The levels of serum E2 and AMH in α group > γ group > β group, and the levels of serum FSH and LH in β group > γ group > α group were statistically significant (P<0.05). The growth follicles were α group > γ group > β group, and the atresia follicles were β group > γ group > α group, and there was a statistical difference among all groups (P<0.05). There was no difference in luteal number among the three groups (P>0.05). In terms of CYR61 and CTGF protein expression, α group > γ group > β group, and in terms of caspase-3, β group > γ group > α group had statistical significance (P<0.05). It is concluded that intervention with umbilical cord mesenchymal stem cells can significantly improve the expression levels of CYR61 and AMH, reduce the level of FSH, promote cell survival, improve the reproductive quality of mice, and restore the physiological function of the ovary. It is feasible to treat premature ovarian failure with umbilical cord mesenchymal stem cells.

MSC Transplantation in Eight Severe COVID-19 Patients: Can Cytokine Storm Be Reversed?

The COVID-19 disease is a global pandemic, with the first case diagnosed in December 2019, as reported by World Health Organization (WHO). In 17% of patients, COVID-19 causes severe Acute Respiratory Distress Syndrome (ARDS) due to release of large amounts of pro-inflammatory cytokines and chemokines in the lungs. In a retrospective observational study from Milan, 9% of the people, who tested positive for COVID-19, needed ICU care with respiratory support. The demand for ICU beds and health care personnel brought significant overload to sustain the care of these patients. Search for effective therapies is underway. However, the result of severe infection of COVID-19 still leads to the inevitable fatalities with the current available therapies. Mesenchymal Stem Cells (MSCs) have been isolated about 30 years ago (Supplementary Data 1). There are over 5,000 articles published on MSCs. Moreover, anti-inflammatory and immunemodulatory properties of MSCs have been well studied. Exogenously administered MSCs are medicinal. They generate positive therapeutic outcomes by secreting bioactive factors that exhibit immunomodulatory, and regenerative effects by fabricating, and secreting antibiotic proteins, where they hone in on sites of injury or disease. Hence, Arnold Caplan has proposed recently to change the name of MSCs to Medicinal Signaling Cells. As MSCs arise from pericytes, they can be isolated from a variety of vascularized tissues. Each separate tissue-specific stem cell interacts with its underlying vascular endothelial cells, and adjacent specific pericyte/MSC “Universal Stem Cell Niche” (pMSCs). Each specific pMSCs have both pMSCs common, and unique chemical, and functional features. Meanwhile, the major therapeutic role of pMSCs in vivo at various sites of disease or injury are very similar when comparing these different pMSCs. Over the past decade, the emphasis has shifted toward harnessing the pMSCs’ ability to produce factors and cytokines that stimulate innate tissue repair, modulate inflammation, and immune responses. MSCs express function on Toll-Like Receptors (TLRs). Triggering different TLRs, depending on exposure times promote either pro- or anti-inflammatory function in MSCs. Pre-clinical studies demonstrated that the majority of infused MSCs initially distributed in the lungs. Subsequent studies showed improved pulmonary functions beginning shortly after administration with no evidence of pulmonary safety risk. These studies indicated the local beneficial MSCs-mediated effect on pulmonary airways . A recent pilot study from China explored the therapeutic outcomes of MSC transplantation in seven poor prognoses COVID-19 patients with pneumonia. The results revealed that MSC transplantation was safe and effective treatment option. The peripheral lymphocytes increased after the treatment, and the overactivated cytokine-secreting immune cells disappeared in 3-6 days. A group of regulatory DC cell population dramatically increased. Meanwhile, the level of TNF-α is significantly decreased, while IL-10 increased in the MSC transplantation group compared to the placebo control group. Furthermore, the gene expression profile showed MSCs were ACE2- and TMPRSS2-, which indicated MSCs were free from COVID-19 infection . Here we report our clinical observations of eight cases, before and after MSC transplantation, to assess the clinical therapeutic effects of MSC transplantation on COVID-19 severe/critically severe patients. Though broader studies are needed, we advised a clinical application protocol and algorithm by evaluating the poor prognostic markers significantly related by MSC transplantation to prevent the overload in ICU clinics, as well as, to shorten hospitalization time.

Repairing effect of Schwann cells combined with mesenchymal stem cells on optic nerve injury in rats.

To investigate the effect and the underlying mechanisms of combined transplantation of Schwann cells (Scs) and Mesenchymal stem cells (MSCs) on optic nerve injury in rats. A total of 160 normal healthy adult rats were randomly divided into 4 groups: optic nerve injury group, optic nerve injury + Sc transplantation group, optic nerve injury + MSC transplantation group and optic nerve injury + Sc + MSC transplantation group. The optic nerve in the left eye of each rat was damaged via clamping to establish a model of optic nerve injury, and the right eye was used as self-control. Scs + MSCs, Scs alone, MSCs alone and normal saline were injected into the vitreous space, respectively. After the treatment, the optic nerve tissues were collected and subjected to hematoxylin-eosin (HE) staining. Next, the morphologic and pathological changes of rats in each group were observed. Retrograde labeling was utilized to count the number of retinal ganglion cells (RGCs) in the optic nerve tissues. The apoptosis of RGCs was detected using flow cytometry. Western blot was carried out to measure the protein expression level of B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax). The expression and distribution of brain-derived neurotrophic factor (BDNF) and growth associated protein-43 (GAP-43) in the optic nerve of rats in each group were detected by immunohistochemistry. Transplantation of Scs and MSCs could maintain the morphological structures of the retina and optic nerve of rats, increase the amount of RGCs in optic nerve tissues, reduce the apoptosis of RGCs, promote the expression of the Bcl-2 protein and decrease the expression of Bax protein. In addition, our joint transplantation strategy also showed an important role in repairing optic nerve injury by clearly promoting the secretion and expression of BDNF and GAP-43, which indicated a better curative effect than that of separate application of Scs or MSCs. Therapy with combined use of Scs and MSCs has a significant therapeutic effect in repairing optic nerve injury.

The effect of bone marrow mesenchymal stem cell transplantation on hypoxic pulmonary hypertension in rats

Objective: To study the influence of bone marrow mesenchymal stem cells (MSCs) transplantation on hypoxic pulmonary hypertension (HPH) in rats. Methods: MSCs in SD rats were separated, cultivated, identified in vitro, and labeled by the green fluorescence protein (GFP) adenovirus. Healthy male SD rats were randomly divided into four groups: normal control group (NC group) and HPH group, with eight rats in each group respectively; HPH + mesenchymal stem cell transplantation group (MSCs group) and HPH + vascular endothelial growth factor + mesenchymal stem cell transplantation group (VEGF + MSCs group), with twenty-four rats in each group respectively. In this experiment, intermittent normobaric hypoxia was employed to establish the pulmonary hypertension rat models, with stem cells transfected and transplanted. The mean pulmonary artery pressure (mPAP) was observed in rats to calculate the right ventricular hypertrophy index (RVHI); the morphological changes of pulmonary arterioles in each group of rats were observed under the microscope; the distribution and manifestation of MSCs fluorescently labeled by adenovirus transfection were observed in pulmonary arterioles under the fluorescence microscope at the set time points of 7 d, 14 d and 28 d after the transplantation of stem cells. Results: For NC group, the mPAP (mmHg) was 15.5 ± 1.5 at 28 d, while the mPAP in HPH, MSCs and VEGF + MSCs groups were 26.1 ± 1.9, 21.6 ± 2.7 and 20.1 ± 2.9 respectively which were apparently higher than that in NC group ( p < .01). Compared with HPH group ( p < .01), the mPAP was obviously decreased in MSCs and VEGF + MSCs groups. There was no significant difference between MSCs and VEGF + MSCs groups. At 28 d, RVHI for NC group was 0.28 ± 0.02, while the RVHI in HPH, MSCs and VEGF + MSCs groups were 0.43 ± 0.07, 0.34 ± 0.03 and 0.35 ± 0.01 respectively which were apparently higher than that in NC group ( p < .01). In comparison with HPH group, RVHI was significantly decreased in MSCs and VEGF + MSCs groups ( p < .05). There was no significant difference between MSCs and VEGF + MSCs groups. For HPH group, at 28 d, pulmonary arterioles were apparently thickened, with luminal stenosis & obliteration and incomplete endothelial cells. Compared with HPH group, pulmonary arterioles in MSCs group became thinning, with the lumen unobstructed and the integrity of endothelial cells improved. The changes in the manifestation of MSCs and VEGF + MSCs groups were not significant. Conclusions: The transplantation of MSCs can improve the remodeling of pulmonary arterioles to partially reverse the progress of HPH; the combined transplantation of VEGF and MSCs doesn’t improve the effect of MSC transplantation.

Serum IFN-\u03b3 levels predict the therapeutic effect of mesenchymal stem cell transplantation in active rheumatoid arthritis

BackgroundTo explore the mechanism of the different clinical efficacies of mesenchymal stem cell transplantation (MSCT) and identify a possible serum biomarker for predicting the therapeutic effect of MSCT in rheumatoid arthritis (RA) patients.MethodsA total of 105 patients with persistently active RA and poor responses to traditional medication were randomly divided into MSCT and control groups. Outcomes were evaluated according to the 28-joint Disease Activity Score and Health Assessment Questionnaire, serological indicators, regulatory T cell (Treg) to T helper 17 (Th17) cell ratio, and inflammatory cytokine levels. Twelve weeks after MSCT, the outcomes of the MSCT group were evaluated according to the European League against Rheumatism response criteria. Patients with a good or moderate response were added to the response group, and those with no response were added to the no-response group.ResultsNo serious adverse events were reported for either MSCT subgroup (28 in the response group and 24 in the no-response group). The therapeutic effects lasted for 48 weeks without continuous administration. Notably, a transient increase in serum IFN-γ (>2 pg/ml) levels was observed in the response group, but not in the no-response group. Furthermore, an increase in IL-10 levels and the Treg/Th17 ratio and a reduction in IL-6 levels appeared 2–3 weeks after the transient IFN-γ increase.ConclusionsAllogeneic MSCT is safe and feasible, and we propose high serum IFN-γ levels as a potent biomarker for predicting MSCT response.Trial registration chictr.org, ChiCTR-ONC-16008770. Registered 3 July 2016, http://www.chictr.org.cn/showproj.aspx?proj=14820

Evaluation of Left Ventricular Regional Systolic Function Using Tissue Doppler Echocardiography After Mesenchymal Stem Cell Transplantation in Rabbits With Myocardial Infarction.

The aim of this work was to assess left ventricular (LV) regional systolic function in rabbits with myocardial infarction after allogeneic mesenchymal stem cell (MSC) transplantation using quantitative tissue velocity imaging. Thirty New Zealand White rabbits were assigned into 3 groups randomly: a sham-operated group (n = 10), a myocardial infarction (MI) group (n = 10), and a MSC transplantation group (n = 10). Mesenchymal stem cells (1 × 10(7) in total) were delivered into 5 spots around the left anterior descending artery (LAD) blood supply area via direct intramyocardial injections 1 hour after LAD ligation in the MSC group, whereas the MI group received the same amount of phosphate-buffered saline injections. Echocardiography was performed before LAD ligation and 1 day and 2 weeks after MSC transplantation, respectively. The peak systolic velocity (Vs) of each LV wall segment was measured. The myocardial slices were harvested for histologic staining after the last echocardiographic examination. The velocity curves for the LV myocardium before LAD ligation had a trend showing that the Vs value decreased gradually from basal to apical segments. The Vs values for the LV segments around the infarcted area in the MSC group decreased significantly compared with the sham group (P < .05) 1 day after MSC transplantation, whereas they increased significantly 2 weeks after MSC transplantation compared with 1 day after LAD ligation (P < .05). This study demonstrates that quantitative tissue velocity imaging may provide a promising approach to quantitatively assessing LV regional systolic function before and after MSC transplantation.

Expression of Wnt and Notch signaling pathways in inflammatory bowel disease treated with mesenchymal stem cell transplantation: evaluation in a rat model.

IntroductionThe purpose of this study was to investigate the expression of Wnt and Notch signaling pathway-related genes in inflammatory bowel disease (IBD) treated with mesenchymal stem cell transplantation (MSCT).MethodsTNBS (2,4,6-trinitrobenzene sulfonic acid) was used to establish IBD in a rat model. Mesenchymal stem cells (MSCs) were transplanted via tail vein transfusion. Saline water was used in a control group. The expression of Wnt and Notch main signaling molecules was screened by gene chips and verified by quantitative reverse transcription-polymerase chain reaction in the IBD rat model on day 14 and day 28 after transplantation.ResultsThe IBD rat models were successfully established and MSCs were transplanted into those models. Genome-wide expression profile chips identified a total of 388 differentially expressive genes, of which 191 were upregulated and 197 were downregulated in the MSC-transplanted group in comparison with the IBD control group. Real-time quantitative polymerase chain reaction results showed that the level of Olfm4 mRNA expression in the IBD group (2.54±0.20) was significantly increased compared with the MSCT group (1.39±0.54) and the normal group (1.62±0.25) (P <0.05). The Wnt3a mRNA was more highly expressed in IBD rats (2.92±0.94) and decreased in MSCT rats (0.17±0.63, P <0.05). The expression of GSK-3β mRNA was decreased in the setting of inflammation (0.65±0.04 versus 1.00±0.01 in normal group, P <0.05) but returned to normal levels after MSCT (0.81±0.17). The expression of β-catenin was observed to increase in IBD tissues (1.76±0.44) compared with normal tissues (1.00±0.01, P <0.05), but no difference was found in the MSCT group (1.12±0.36). Wnt11 declined at 14 days and returned to normal levels at 28 days in the IBD group; in comparison, a significantly lower expression was found in MSCT rats. There were no differences in the expression of Fzd3, c-myc, TCF4, and Wnt5a in inflammation, but all of those genes declined after MSCT treatment.ConclusionsThe canonical Wnt and Notch signaling pathways are activated in IBD and may be suppressed by stem cell transplantation to differentiate into intestinal epithelium after MSCT. Moreover, the non-canonical Wnt signaling may be inhibited by canonical Wnt signaling in the setting of inflammation and may also be suppressed by MSCT.

Mesenchymal stem cells secrete brain-derived neurotrophic factor and promote retinal ganglion cell survival after traumatic optic neuropathy.

The study aimed to investigate whether intravitreal injection of mesenchymal stem cells (MSCs) cultivated in vitro could increase the number of survived retinal ganglion cells (RGCs) after traumatic optic neuropathy and sought to identify potential mechanisms underlying such growth. The right eye of 24 cats in the MSC transplantation group accepted intravitreal injection of MSCs, and the other 24 cats in the phosphate buffer (PBS) control group received isotonic saline after traumatic optic neuropathy. The RGCs' survival rate in separated retinal and brain-derived neurotrophic factor (BDNF) expression were observed by Dil labeling and Enzyme-Linked Immuno Sorbent Assay (ELISA), respectively, at 3, 7, 14, and 28 days after transplantation. Quantitative analysis showed that RGCs were significantly attenuated at 3, 7, 14, and 28 days after transplantation in both areas of the retina (peripheral, P7d = 0.0011, P14d < 0.001, P28d < 0.001; central, P3d = 0.0437, P7d = 0.0067, P14d < 0.001, P28d < 0.001). Mean density of RGCs in the MSC transplantation group was significantly higher than that of the PBS control group after 14 days of treatment (P < 0.001). The homogenates BDNF (hBDNF) in the MSC transplantation group was obviously higher than that in the PBS control group at 14 and 28 days (P < 0.05). The MSCs transplanted into the retina of cats can slow down RGC apoptosis and steadily express BDNF. The MSC-mediated neuroprotection after optic nerve injury may be related to BNDF.

A Transgenic Tri-Modality Reporter Mouse

Transgenic mouse with a stably integrated reporter gene(s) can be a valuable resource for obtaining uniformly labeled stem cells, tissues, and organs for various applications. We have generated a transgenic mouse model that ubiquitously expresses a tri-fusion reporter gene (fluc2-tdTomato-ttk) driven by a constitutive chicken β-actin promoter. This “Tri-Modality Reporter Mouse” system allows one to isolate most cells from this donor mouse and image them for bioluminescent (fluc2), fluorescent (tdTomato), and positron emission tomography (PET) (ttk) modalities. Transgenic colonies with different levels of tri-fusion reporter gene expression showed a linear correlation between all three-reporter proteins (R2=0.89 for TdTomato vs Fluc, R2=0.94 for Fluc vs TTK, R2=0.89 for TdTomato vs TTK) in vitro from tissue lysates and in vivo by optical and PET imaging. Mesenchymal stem cells (MSCs) isolated from this transgenics showed high level of reporter gene expression, which linearly correlated with the cell numbers (R2=0.99 for bioluminescence imaging (BLI)). Both BLI (R2=0.93) and micro-PET (R2=0.94) imaging of the subcutaneous implants of Tri-Modality Reporter Mouse derived MSCs in nude mice showed linear correlation with the cell numbers and across different imaging modalities (R2=0.97). Serial imaging of MSCs transplanted to mice with acute myocardial infarction (MI) by intramyocardial injection exhibited significantly higher signals in MI heart at days 2, 3, 4, and 7 (p<0.01). MSCs transplanted to the ischemic hindlimb of nude mice showed significantly higher BLI and PET signals in the first 2 weeks that dropped by 4th week due to poor cell survival. However, laser Doppler perfusion imaging revealed that blood circulation in the ischemic limb was significantly improved in the MSCs transplantation group compared with the control group. In summary, this mouse can be used as a source of donor cells and organs in various research areas such as stem cell research, tissue engineering research, and organ transplantation.

A novel treatment regimen for acute liver failure based on a combination of mesenchymal stem cells transplantation and IL-lRa-loaded chitosan nanoparticles

To evaluate whether a combination therapy using allogeneic mesenchymal stem cell (MSC) transplantation and interleukin-1 receptor antagonist (IL-1Ra) chitosan nanoparticles is more robust than MSC transplantation alone for treating acute liver failure and to investigate the mechanisms of the improved therapeutic effect using a swine model system. IL-1Ra-loaded nanoparticles were made of lactosylated chitosan-FITC using the electrostatic spray method and analyzed by enzyme-linked immunosorbent assay. The active live targeting of these nanopaticles were investigated by fluorescence microscope and flow cytometer(FCM). The combined therapy was given and the Detailed Steps as follows: Chinese experimental mini swine were given D-galactosamine to build models of acute liver failure. Thirty-four pigs were randomly divided into five groups. In the control group(A),the normal saline was injected into liver via portal veins after 24 h; in IL-1Ra group(B), IL-1Ra was injected via ear veins 6 h before normal saline; In the MSCs transplantation group (C), 8 * 107 MSCs were injected into liver via portal veins after 24 h; IL-1Ra together with MSCs transplantation group(D) and nanopaticles group(E) as follows: on the one hand, 8 * 107 MSCs were injected into liver via portal veins after 24 h, on the other hand, rhIL-1Ra in group C or IL-1Ra chitosan nanopaticles in group D was injected via ear veins respectively at 6 h before. Liver function, serum inflammation and pathological changes were measured. The fate of MSCs was also observed. The profiles in vitro shown that there was a steady-state release after a fast linear release; The live targeting of the lactosylated chitosan-based nanoparticles was achieved by ligand-receptor specificity; The biochemical assay, the serum inflammation lever and pathological changes of the nanopaticles group were all greatly different from the other groups, the hepatocytes grow rate was significantly improved after 1 week; The liver engraftment was very low in group C and D with significantly higher numbers found in nanopaticles group, but the differentiation of MSCs after 2 weeks relatively rare; western blot showed that there was more HGF and VEGF secreted in nanopaticles group. IL-lRa-loaded lactosylated chitosan-based nanopaticles have significant liver targeting abilities and slow release characteristics in PBS solution. The combined therapy showed great synergistic effects through suppression of liver inflammation and paracrine.

Effects of intraperitoneal injection of marrow mesenchymal stem cells on intestinal barrier in acute pancreatitis

To explore the effects of intraperitoneal injection of mesenchymal stem cells (MSC) on intestinal barrier in serve acute pancreatitis (SAP) rats. MSC were harvested and cultured from femurs of one male SD rat. And 30 female SD rats were divided into 3 groups: control group (n = 6), SAP group (n = 12) and MSC transplantation group (n = 12). SAP was induced by intraperitoneal injection of L-arginine (2 g/kg) twice in SAP and MSC groups. In MSC group, the third-generation MSC (5×10(6)) were injected intraperitoneally once daily for 3 days. All rats were sacrificed after 72 h. The histomorphologic alternations of small intestine were measured to evaluate the therapeutic effect of MSC transplantation. Reverse transcription (RT)-PCR were used to identify the expression of TNF-α mRNA and IL-1β mRNA in small intestine and pancreas. Small intestine and pancreatic samples were examined for the engraftment of donor-derived MSC by Y chromosome in situ hybridization analysis. Compared with SAP group, histomorphologic alternations of small intestine significantly lower in MSC group (4.17 ± 0.28 vs 3.00 ± 0.33, P < 0.05). The relative expression quantity of TNF-α mRNA and IL-1β mRNA in pancreas were both significant higher in SAP and MSC groups than those in control group (3.10 ± 0.73 and 1.92 ± 0.37 vs 0.51 ± 0.24, 4.60 ± 0.59 and 2.43 ± 0.39 vs 1.15 ± 0.18, all P < 0.05). Compared with SAP group, the expression quantity of TNF-α mRNA and IL-1β mRNA in pancreas significantly lower in MSC group (both P < 0.05). The relative expression quantity of TNF-α mRNA and IL-1β mRNA in small intestine were both significant higher in SAP and MSC groups than those in control group (2.73 ± 0.91 and 1.55 ± 0.48 vs 0.62 ± 0.20, 5.20 ± 0.94 and 2.10 ± 0.34 vs 0.99 ± 0.10, all P < 0.05). The expressions of TNF-α mRNA and IL-1β mRNA in MSC group were lower than those in SAP group (both P < 0.05). Sry gene was not detected in pancreatic and intestinal tissue of MSC-treated rats. Syngraft MSC exert protective effects on pancreas and small intestine injury. And their beneficial effects are primarily mediated via indirect actions but not by their differentiation into target cells.

The comparison of curative effects between mesenchymal stem cells of the human umbilical cord and mesenchymal stem cells of rat bone marrow in rats with spinal cord injuries

Objective To observe changes in the functional recovery and immunological rejection of rats after spinal cord injuries with the treatment of transplanting human umbilical cord mesenchymal stem cells and rat bone marrow mesenchymal stem cells,to find the right stem cells for clinical procedures through this experiment,and to investigate the mechanism of immunological rejection after transplanting the cells.Methods Forty SD rats were selected randomly and divided into 3 groups.16 rats were placed in the umbilical cord mesenchymal stem cell transplantation group,16 rats in the bone marrow mesenchymal stem cell transplantation group,and 8 rats in the PBS group.The levels of 1I-2 and IL-10 in the blood of each group were detected at different times (1W and 2M,and 3 months after the cell transplantation) to reflect the systemic immunological rejection of the goat after transplantation.The CD3 + cells in the cell transplantation areas of each group were also detected with immunohistochemistry to reflect local immune rejection after transplantations.Results 1.The level of IL-2 was obviously higher than the PBS group lW and 2M and 3 months after the cell transplantation (P ＜0.05).Meanwhile,IL-10 was Contrary to the IL -2.2.The level of IL-2 was obviously higher than the PBS group 1W and 2M and 3 months after the cell transplantation (P ＜ 0.05).Meanwhile,IL-10 was Contrary to the IL-2.3.The quantity of CD3+cells in all groups were much at the acute period (1 W after cell transplantation),but only the quantity of CD3+cells in the bone marrow mesenchymal stem cell transplantation group was much larger compared to the other groups at the chronic period 3 months after cell transplantation (P ＜ 0.05).Conclusion Systemic and local immunological rejections at acute or chronic periods will appear in rats with spinal cord injuries that underwent stem cell transplantations.Compared with bone marrow mesenchymal stem cells,the umbilical cord mesenchymal stem cells significantly have lower immunogenicity and more suited for functional recoverv. Key words: Spinal cord injury; Umbilical cord mesenchymal stem cells; Bone marrow mesenchymal stem cells; Immunological rejection; Functional recovery

Angiogenesis and bone regeneration by allogeneic mesenchymal stem cell intravenous transplantation in rabbit model of avascular necrotic femoral head

AimTo explore the feasibility of allogeneic mesenchymal stem cells (MSCs) transplanted intravenously for angiogenesis and bone repair in a rabbit model of avascular necrosis of femoral head (ANFH). Materials and methodsForty-five rabbits were randomized into three groups: a blank control group (without treatment), a necrotic control group (ANFH induced but without therapy), and an MSC transplantation group (ANFH induced and treated with MSC transplantation). The biopsies, blood sampling, and imaging examinations were performed on each animal at different time points (2, 4, and 6 wk). To monitor angiogenesis and bone repair progress, examinations included real-time polymerase chain reaction, Western blot analysis, x-ray, computed tomography, Masson trichrome staining, picrosirius red staining, and immunohistochemical staining. ResultsNecrosis and bone collapse were observed in bilateral femoral heads of necrotic rabbits of the necrotic control group, whereas the femoral head morphology was generally restored in the MSC transplantation group. The mRNA levels of Cbfa1, BMP, VEGF, and OPN in bone tissue were significantly higher in the MSC transplantation group than in the necrotic control group. In addition, the total protein amount of Cbfa1 in the MSC transplantation group was also significantly higher than that in the necrotic control group (P < 0.05). ConclusionIntravenous transplantation of allogeneic MSCs can promote vascular and bone regeneration in the necrotic region of the femoral head in a rabbit model of ANFH. The results of our study suggest that the intravenous transplantation of MSCs could be a potential and minimally invasive treatment option for ANFH patients.

Effect of mesenchymal stem cells on cardiac function and restenosis of injured artery after myocardial infarction

Although earlier studies have shown that the transplantation of mesenchymal stem cells (MSCs) might improve cardiac functions after myocardial infraction, its role on vascular restenosis after percutaneous coronary intervention (PCI) remains controversial. The aim of this study was to investigate the effects of MSCs on the restenosis of injured artery following balloon angioplasty in a rabbit model with both myocardial infarction reperfusion and atherosclerotic stenosis carotid artery by balloon injury. After the animal model was established for myocardial infraction reperfusion and atherosclerotic stenosis carotid artery by balloon injury, the rabbits received an intravenous transplantation of MSCs. And an equal volume of phosphate buffered solution was administered for the control group. The animal vascular tissue and myocardium tissue were excised at different time points post-transplantation and used to detect the homing of MSCs and the expressions of platelet-endothelial cell adhesion molecule-1 (CD31) and proliferating cell nuclear antigen (PCNA) by immunohistochemical staining. Four weeks later, vascular restenosis was analyzed by angiography of bilateral carotid arteries and the vascular tissues were used for histological studies. At one week post-transplantation, the 4',6-diamidino-2-phenylindole (DAPI)-labeled MSCs could be detected in myocardial infarction and injured intima. And the intimal expression of CD31 was observed at 2 weeks in the MSCs transplantation group. Yet the expression of PCNA was significantly lower in the MSCs transplantation group than that in the control group (50.5% ± 3.6% vs 23.4% ± 2.8%, P < 0.05). At 4 week post-transplantation, the neointimal area of injured vessels and the vascular restenosis were significantly lower in the MSCs transplantation group than those in the control group (0.092 ± 0.009 vs 0.189 ± 0.007, P < 0.05; 41.7 ± 3.7 vs 61.3 ± 1.6, P < 0.05). Furthermore the MSCs transplantation group demonstrated improved cardiac functions, reduced myocardial infarct size (21.7% ± 2.2% vs 34.3% ± 1.8%, P < 0.05) and significantly increased capillary density around infarction foci (33.6% ± 2.1% vs 20.8% ± 2.6%, P < 0.05) versus the control group. The transplantation of MSCs plays significant roles in cardiac repairing in terms of improved cardiac functions, accelerated repair of injured vessels, suppression of neointimal hyperplasia and reduced restenosis of injured vessels.

Rosuvastatin Treatment Activates JAK-STAT Pathway and Increases Efficacy of Allogeneic Mesenchymal Stem Cell Transplantation in Infarcted Hearts

Widespread death of implanted cells hampers the development of stem cell therapy for acute myocardial infarction (AMI). Our previous studies indicated that statins can protect implanted mesenchymal stem cells (MSCs) against the post-infarct microenvironment, thus increasing the therapeutic effect. However, the underlying mechanisms are unclear. The JAK-STAT pathway participates in regulation of stress responses of the myocardium to various insults. This study aimed to detect whether rosuvastatin (ROSU) facilitates the survival, engraftment, and differentiation of allogeneic bone marrow-derived MSCs in the post-infarct heart via the JAK-STAT signaling pathway. Female Sprague-Dawley rats were randomized into 5 groups: AMI (control), ROSU gavage (group R), MSCs transplantation (group M), MSCs and ROSU (group M+R), or MSCs, ROSU and a JAK2 inhibitor AG-490 (group M+R+AG). MSCs from male rats were injected into the myocardium 1 week after AMI. Cardiac function and histology, as well as the expression of Y-chromosomal genes and JAK-STAT signaling proteins, were examined at 4 weeks after transplantation. Better functional recovery, increased survival and differentiation of MSCs occurred in group M+R. Furthermore, phosphorylation of JAK2 and STAT3 was higher in group M+R. The effects of ROSU, as well as of activated JAK-STAT proteins, could be attenuated by AG-490. ROSU treatment improves the efficacy of stem cell transplantation in infarcted hearts by activation of the JAK2-STAT3 signaling pathway.

Paracrine Role for Mesenchymal Stem Cells in Acute Myocardial Infarction

The differentiation function and paracrine action of mesenchymal stem cells (MSCs) have been reported previously, although it is not clear which accounts for the short-term effects on post-MSC transplantation. In this study, rats with acute myocardial infarction (AMI) induced by occlusion of the left coronary artery were randomized to the MSC transplantation group and MI group. The differentiation and paracrine action of MSC were observed after 2 weeks of MI. We found that MSC transplantation improved cardiac function [left ventricular end-systolic pressure (LVESP), left ventricular end-diastolic pressure (LVEDP), left ventricular +dp/dt and ejection fraction (EF)]. Only a few grafted MSC differentiated into myocardial cells or vascular endothelial cells. Conversely, a significant increase in capillary density was observed. MSC transplantation upregulated the expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). These results indicate that an increase in capillary density and improved cardiac function post-MSC transplantation are closely related to the up-regulation of VEGF and bFGF.