Umbilical Cord Blood Mesenchymal Stem Cells Research Articles

Umbilical cord blood-mesenchymal stem cells and carvedilol reduce doxorubicin- induced cardiotoxicity: Possible role of insulin-like growth factor-1.

In this study, we tried to demonstrate the effects of adding human umbilical cord blood mesenchymal stem cells (hUCB-MSCs) to carvedilol in improving the doxorubicin- induced cardiotoxicity in rats. Rats were randomly divided into four groups: group 1: control group, group 2: doxorubicin untreated group, group 3: rats injected with doxorubicin and received carvedilol, and group 4: rats injected with doxorubicin and received carvedilol and stem cell-treated. Electrocardiography (ECG) was performed to assess cardiac function after animals were sacrificed. Cardiac muscle sections were examined histologically using H&E, Masson trichrome and immunohistochemically using caspase 3 immunostaining. The morphometric and statistical analysis was performed. Levels of malondialdehyde (MDA), superoxide dismutase (SOD), insulin-like growth factor (IGF-1), and vascular endothelial growth factor (VEGF) were measured. We concluded that combination of hUCB-MSCs and carvedilol markedly improves histological and immunohistochemical structure of cardiac muscle fibers and restores cardiac function in doxorubicin- induced cardiotoxicity in rats.

Effects of mesenchymal stem cells on pulmonary fibrosis

Objective To investigate the effects of mesenchymal stem cells (MSCs) on pulmonary fibrosis under different conditions (different types, different doses and different transplantation times in the same dose). Methods Seventy-two male SD rats were equally divided into 6 groups by random number table method: Group A received intratracheal instillation of NS (0.1 ml). Group B, C, D, E and F received intratracheal instillation of bleomycin (5 mg/kg in 0.1 ml NS). On the 1st day after bleomycin administration, Group A and B received NS (1.0 ml) via the tail vein, while Group C received bone marrow mesenchymal stem cells (BMSCs) 1×106, Group D received umbilical cord blood mesenchymal stem cells (UCB-MSCs) 1×106, Group E received BMSCs 5×105 and Group F received BMSCs 1×107 via the tail vein, respectively. On the 8th day after bleomycin administration, Group E received BMSCs 5×105 again. Half of rats in each group were sacrificed respectively on the 28th and 42nd day after bleomycin administration. The pathologic changes, transforming growth factor-β1 (TGF-β1), hydroxyproline (HYP), matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-1 (TIMP-1) of the lung tissues were investigated. Results Alveolar structure was severely in disorder, and massive collagenous fiber was deposited in Group B. In Group B, C, D, E, F and A, the scores of pulmonary fibrosis were decreased in sequence [(3.00±0.00), (2.17±0.75), (1.60±0.89), (1.33±0.52), (1.00±0.00), (0.00±0.00) on the 28th day after bleomycin administration, and (3.00±0.00), (2.40±0.55), (1.75±0.96), (1.50±0.84), (-), (0.00±0.00) on the 42nd day after bleomycin administration], while the levels of TGF-β1, HYP and MMP-2/TIMP-1 were respectively decreased in sequence as above. The levels of TGF-β1, HYP and MMP-2/TIMP-1 were positively associated with pulmonary alveolitis and fibrosis scores respectively (P=0.000). Conclusion MSCs transplantation can suspend the progress of pulmonary fibrosis, and the mechanism may be related to reducing the level of TGF-β1 and improving the imbalance of MMP/TIMP. Choosing UCB-MSCs or multiple transplantation may improve the therapeutic effects of pulmonary fibrosis. Key words: Pulmonary fibrosis; Bleomycin; Bone marrow mesenchymal stem cells; Umbilical cord blood mesenchymal stem cells

Isolation and characterization of in vitro culture of hair follicle cells differentiated from umbilical cord blood mesenchymal stem cells.

The present investigation explored the in vitro culture, isolation and characterization of hair follicle cell differentiation from umbilical cord blood mesenchymal stem cells (MSCs). Flow cytometry was used to obtain MSCs from the isolation and purification of human umbilical cord blood MSCs. Culture suspension of hair follicle organ was centrifuged and the supernatant used in the culture medium of MSCs, and the entire process of induced differentiation was recorded by photomicroscopy. The expression level of surface marker CK15 of hair follicle cells obtained from induced differentiation was detected with immunofluorescence. RT-PCR method was used to further detect the difference in expression of CK15 between hair follicle cells and umbilical cord blood MSCs, and statistical analysis was carried out. CD44+CD29+ double-labeled cells accounted for 50.8% of all the samples of umbilical cord blood MSCs in this study. The diameter of hair follicle cells differentiated from umbilical cord blood stem cells reached 800×10−3 mm after 3 weeks of cell culture. Based on the detection and colocalization of CK15 expression in induced hair follicle cells, the overlap ratio between CK15 and nuclei reached 83% in hair follicle cells, which was obviously higher than that in umbilical cord blood stem cells. The difference had statistical significance (P<0.05). In conclusion, hair follicle cells can be successfully differentiated from umbilical cord blood stem cells by using the supernatant from hair follicle cells. This method can be used for high-speed induced differentiation with high purity, which is promising for clinical application.

Metabolomics Analysis of the Osteogenic Differentiation of Umbilical Cord Blood Mesenchymal Stem Cells Reveals Differential Sensitivity to Osteogenic Agents.

Mesenchymal stem cells (MSCs) of fetal origin, such as umbilical cord blood MSCs (UCB MSCs), have emerged as a promising cell source for musculoskeletal tissue regeneration because of their higher proliferation potential, lack of donor site morbidity, and their off-the-shelf potential. MSCs differentiated toward the osteogenic lineage exhibit a specific metabolic phenotype characterized by reliance to oxidative phosphorylation for energy production and reduced glycolytic rates. Currently, limited information exists on the metabolic transitions at different stages of the osteogenic process after osteoinduction with different agents. Herein, the osteoinduction efficiency of BMP-2 and dexamethasone on UCB MSCs was assessed using gas chromatography–mass spectrometry (GC-MS) metabolomics analysis, revealing metabolic discrepancies at 7, 14, and 21 days of induction. Whereas both agents when administered individually were able to induce collagen I, osteocalcin, and osteonectin expression, BMP-2 was less effective than dexamethasone in promoting alkaline phosphatase expression. The metabolomics analysis revealed that each agent induced distinct metabolic alterations, including changes in amino acid pools, glutaminolysis, one-carbon metabolism, glycolysis, and tricarboxylic acid cycle. Importantly, we showed that in vitro-differentiated UCB MSCs acquire a metabolic physiology similar to primary osteoblasts when induced with dexamethasone but not with BMP-2, highlighting the fact that metabolomics analysis is sensitive enough to reveal potential differences in the osteogenic efficiency and can be used as a quality control assay for evaluating the osteogenic process.

344 - Comparison of undifferentiated vs chondrogenic predifferentiated human umbilical cord-blood mesenchymal stem cells for cartilage repair in a rat model

344 - Comparison of undifferentiated vs chondrogenic predifferentiated human umbilical cord-blood mesenchymal stem cells for cartilage repair in a rat model

Human umbilical cord blood-mesenchymal stem cells transplantation renovates the ovarian surface epithelium in a rat model of premature ovarian failure: Possible direct and indirect effects.

This study aimed to isolate mesenchymal stem cells (MSC) from human umbilical cord blood (HCB) and to explore their influence on the ovarian epithelium after paclitaxel-induced ovarian failure. Ninety-five rats were divided into 6 groups: control, paclitaxel, paclitaxel and saline, HCB-MSC-treated for 2 weeks, HCB-MSC-treated for 4 weeks, and HCB-MSC-treated for 6 weeks. HCB cells were studied for CD34, CD44, and Oct ¾ using flow cytometry. Serum levels of FSH and E2 were measured using ELISA, RT-PCR analysis for human gene; beta-actin (ACTB), immunohistochemical analysis for CK 8/18, TGF-ß, PCNA and CASP-3 were performed. We found that ACTB gene was expressed in all rats' ovaries received HCB-MSC. After 4 weeks of transplantation, there was significant reduction in FSH, elevation in E2 levels, stabilization of the surface epithelium morphostasis, an increase in the antral follicle count and increase in integrated densities (ID) of CK 8/18, TGF-ß, and PCNA expressions and decrease in ID of CASP-3 expression. We concluded that HCB-MSC can restore the ovarian function after paclitaxel injection through a direct triggering effect on the ovarian epithelium and/or indirect enrichment of ovarian niche through regulating tissue expression of CK 8/18, TGF-ß and PCNA. These molecules are crucial in regulating folliculogenesis and suppressing CASP-3-induced apoptosis.

Chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells in vitro.

Different therapeutic techniques have been developed for regeneration of articular cartilage injuries, but none has provided an optimal solution to their treatment. Human umbilical cord blood-mesenchymal Stem Cells (HUCB-MSCs) have been considered as promising alternative cell source for cartilage repair. Examining the success rate of MSCs isolation from HUCB as well as chondrogenic differentiation potential of HUCB-MSCs in vitro. 32 UCB samples were collected, in addition to 5 bone marrow (BM) and 5 peripheral blood (PB) samples, taken as reference controls. Samples were used for mononuclear cells isolation from which MSCs were expanded under complete aseptic conditions, were verified morphologically and through the presence of CD44 and CD105, and absence of CD34. Success rate of UCB-MSCs isolation was (25%), a rate that was lower than those of PB (40%) and BM (80%). Accordingly, certain input parameters have been recommended for successful MSCs isolation from UCB. On selecting samples in which recommended parameters were fulfilled, success rate was increased to 72%. This was together with providing optimal experiment conditions; mainly type of expansion medium, success rate reached 80%. Then, successfully expanded MSCs were subjected to chondrogenic differentiation by culturing in pelleted micromass system in presence of transforming growth factor beta-1 and chondrogenic medium devoid of fetal bovine serum to evaluate their ability to undergo chondrogenesis. Differentiation was verified microscopically using special stains, and proved by reverse transcriptase-polymerase chain reaction for expression of aggrecan and collagen II genes. In conclusion, in vitro differentiation into chondrocytes is possible from HUCB-MSCs.

Immunomodulatory effects of umbilical cord-derived mesenchymal stem cells.

Umbilical cord blood (UCB) is of great interest as a source of stem cells for use in cellular therapies. The immunomodulatory effect of mesenchymal stem cells (MSCs) originating from bone marrow, adipose tissue and amniotic membrane has previously been reported. In this study, MSCs were isolated from UCB with the aim of evaluating their immunomodulatory effects on proliferation of PB lymphocytes by two different techniques; namely, 5-bromo-2-deoxyuridine ELISA and a carboxy fluorescein diacetate succinimidyl ester flow cytometric technique. MSCs were isolated from UCB, propagated until Passage four, and then characterized for cell surface markers by flow cytometry and ability to differentiate towards osteocytes and adipocytes. Immunosuppressive effects on PB lymphocytes were examined by co-culturing mitomycin C-treated UCB MSCs with mitogen-stimulated lymphocytes for 72 hr. Thereafter, proliferation of lymphocytes was detected by CFSE flow cytometry and colorimetric ELISA. The titers of cytokines in cell culture supernatant were also assayed to clarify possible mechanisms of immunomodulation. UCB MSCs suppressed mitogen-stimulated lymphocyte proliferation, which occurs via both cell-cell contact and cytokine secretion. Titers of transforming growth factor beta and IL 10 increased, whereas that of IFN-γ decreased in the supernatants of co-cultures. Thus, UCB MSCs suppress the proliferation of mitogen-stimulated lymphocytes. However further in vivo studies are required to fully evaluate the immunomodulatory effects of UCB MSCs.

The Effect of Umbilical Cord Blood Derived Mesenchymal Stem Cells in Monocrotaline-induced Pulmonary Artery Hypertension Rats.

Pulmonary arterial hypertension (PAH) causes right ventricular failure due to a gradual increase in pulmonary vascular resistance. The purposes of this study were to confirm the engraftment of human umbilical cord blood-mesenchymal stem cells (hUCB-MSCs) placed in the correct place in the lung and research on changes of hemodynamics, pulmonary pathology, immunomodulation and several gene expressions in monocrotaline (MCT)-induced PAH rat models after hUCB-MSCs transfusion. The rats were grouped as follows: the control (C) group; the M group (MCT 60 mg/kg); the U group (hUCB-MSCs transfusion). They received transfusions via the external jugular vein a week after MCT injection. The mean right ventricular pressure (RVP) was significantly reduced in the U group after the 2 week. The indicators of RV hypertrophy were significantly reduced in the U group at week 4. Reduced medial wall thickness in the pulmonary arteriole was noted in the U group at week 4. Reduced number of intra-acinar muscular pulmonary arteries was observed in the U group after 2 week. Protein expressions such as endothelin (ET)-1, endothelin receptor A (ERA), endothelial nitric oxide synthase (eNOS) and matrix metalloproteinase (MMP)-2 significantly decreased at week 4. The decreased levels of ERA, eNOS and MMP-2 immunoreactivity were noted by immnohistochemical staining. After hUCB-MSCs were administered, there were the improvement of RVH and mean RVP. Reductions in several protein expressions and immunomodulation were also detected. It is suggested that hUCB-MSCs may be a promising therapeutic option for PAH.

Oxygen is a critical parameter for chondrogenic differentiation of human umbilical cord blood mesenchymal stem cell in 3D-cultures

Oxygen is a critical parameter for chondrogenic differentiation of human umbilical cord blood mesenchymal stem cell in 3D-cultures

Hearing Restoration with Intravenous Transplantation of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood

Objectives: Confirm the effect of transplantation of human umbilical cord blood mesenchymal stem cells (UCB-MSCs) on hearing restoration in a sensorineural hearing loss (SNHL) animal model. Methods: UCB was collected from pregnant women after obtaining consent, and mesenchymal stem cells (MSCs) were extracted. We established an SNHL model and transplanted UCB-MSCs through the brachial vein of the guinea pigs. The animals were divided into 4 groups: animals with normal hearing, animals with SNHL, animals with SNHL and injected with saline, and animals with SNHL and transplanted with UCB-MSCs. Hearing tests were conducted at 1, 3, and 5 weeks, and the results were compared by grading auditory brainstem response (ABR) recordings and distortion product otoacoustic emissions (DPOAEs) for each treatment. Lastly, cochlear pathological features were examined, and surface preparations and morphological changes in each animal model were compared using hematoxylin and eosin staining and light microscopy studies. Results: In SNHL group, decreased DPOAEs and increased ABR threshold were noted. Furthermore, in the SNHL group, ABR hearing thresholds were unconverted and were similar to those observed in deafness. The transplanted UCB-MSC group showed a significant improvement in hearing threshold (40 dB) compared to that in all the SNHL group (80–90 dB). Examination of the SNHL animals’ cochlear morphological features demonstrated a noticeable lack of spiral ganglion cells and also showed degenerated outer hair cells. However, the transplanted UCB-MSCs showed an increase in spiral ganglion and hair cells. Conclusions: Intravenous transplantation of UCB-MSCs can enhance hearing thresholds, outer-hair cells and increase the number of spiral ganglion neurons (SGNs).

Regenerative Stem Cell Therapy with Umbilical Cord Mesenchymal Stromal Cells in Deaf Animal Model

Objectives: This study was performed to confirm the effect of transplantation of human umbilical cord blood mesenchymal stem cells (UCB-MSCs) on hearing restoration in deaf animal model. Methods: UCB was collected from pregnant women after obtaining consent, and mesenchymal stem cells (MSCs) were extracted. We established deaf animal model and transplanted UCB-MSCs through the brachial vein of the guinea pigs. The animals were divided into 4 groups: animals with normal hearing, animals with sensorineural hearing loss (SNHL), animals with SNHL and injected with saline, and animals with SNHL and transplanted with UCB-MSCs. Hearing tests were conducted at 1, 3, and 5 weeks, and the results were compared by grading auditory brainstem response (ABR) recordings and distortion product otoacoustic emissions (DPOAEs) for each treatment. Lastly, cochlear pathological features were examined. Results: In SNHL group, decreased DPOAEs and increased ABR threshold were noted. And ABR hearing thresholds were unconverted and were similar to those observed in deafness. The transplanted UCB-MSC group showed a significant improvement in hearing threshold (40 dB). Examination of the SNHL animals’ cochlear morphological features demonstrated a noticeable lack of spiral ganglion cells and also showed degenerated outer hair cells. However, the transplanted UCB-MSCs showed an increase in spiral ganglion and hair cells. Conclusions: Intravenous transplantation of UCB-MSCs can enhance hearing thresholds, and regenerate inner ear hair cells and spiral ganglion neurons (SGNs).

Regulation and direction of umbilical cord blood mesenchymal stem cells to adopt neuronal fate

Umbilical cord blood mesenchymal stem cells (UCB-MSCs) transplantation is becoming a promising and attractive cell-based treatment modality for repairing the damaged central nervous system due to its advantages of low immunogenicity, wide range of sources, and less ethical controversy. One of the limitations of this approach is that the proportion of neurons differentiated from UCB-MSCs still remains at low level. Thus, to induce UCB-MSCs to differentiate into neuron-like cells with a higher proportion is one of the key technologies of regenerative medicine and tissue engineering. Many induction protocols with remarkably higher differentiation rate to neurons have been reported. However, each protocol has its pros and cons and whether the neurons differentiated from UCB-MSCs under a certain protocol has normal nerve function remains controversial. Therefore, to guarantee the success of future clinical applications of UCB-MSCs, more investigations should be performed to improve the induction method and differentiation efficiency.

Comparative evaluation of in vivo osteogenic differentiation of fetal and adult mesenchymal stem cell in rat critical-sized femoral defect model

Mesenchymal stem cells (MSCs) can be obtained from various sources. MSCs from different origins appear to have different preferences for differentiation. In this study, we have compared the in vivo osteogenic potential of adult MSCs from adipose tissue (AT) and bone marrow (BM) with fetal MSCs from umbilical cord (UC) and umbilical cord blood (UCB) by using a rat critical-sized femoral defect model. We have also sought to determine whether pretreatment with an osteogenic medium promotes osteogenesis in MSCs. Study groups were divided as follows: (1) defect only, (2) scaffold only, (3) AT MSCs in scaffolds, (4) BM MSCs in scaffolds, (5) UC MSCs in scaffolds and (6) UCB MSCs in scaffolds. Groups with MSCs were further divided with respect to their pretreatment. At 12 weeks after surgery, in vivo osteogenesis was measured radiographically and by micro-computed tomography (CT). Based on quantitative assessment by micro-CT, no significant difference of the mean bone volume fraction value (BV/TV) was seen between adult MSCs (AT and BM MSCs) and fetal MSCs (UC and UCB MSCs). The mean BV/TVs were significantly higher in non-pretreated BM MSC (14.2±1.4%) and UCB MSC (14.0±1.2%) and pretreated UC MSC (14.8±2.0%) than in those with the scaffold only (11.3±1.3%; P<0.05). In addition, AT (from 10.4±1.2% to 13.1±2.2%) and UC (from 10.3±0.7% to 14.8±2.0%) MSCs from solid tissues showed a significant increase in the mean BV/TV with pretreatment (P<0.05). In contrast, BM MSC (from 14.2±1.4% to 10.9±1.2%) and UCB MSC (from 14.0±1.2% to 11.6±1.0%) from non-solid tissues showed a significant decrease with pretreatment (P<0.05).

Study of inhibiting and killing effects of transgenic LIGHT human umbilical cord blood mesenchymal stem cells on stomach cancer

To study the inhibition and killing effect of transgenic LIGHT umbilical cord blood mesenchymal stem cells (UCBMSCs) on stomach carcinoma. The LIGHT gene was recombined to construct the transfer plasmid pGC-FU-LIGHT by infusion technique. The 293T cells were co-transfected with the transfer plasmid pGC-FU-LIGHT, the construction plasmid Helper 1.0 and the envelope plasmid Helper 2.0 with the help of lipofectamine 2000 to produce lentiviral particles. Transgenic UCBMSCs(MSC-LIGHT) and empty carrier UCBMSCs (MSC) were obtained. Human gastric cancer cell SGC-7901 was injected into nude mice subcutaneously groin. The model of transplanted human gastric cancer cell SGC-7901 in nude mice was established. Tumorigenesis nude mice were separated into three groups randomly with 5 in each group: MSC-LIGHT group, MSC group, and NS group. Three groups of nude mice were injected around the tumor with MSC-LIGHT, MSC and NS every other day for 3 times. Four weeks later, the transplanted gastric cancer volume was measured. The expressions of LIGHT in the three groups were determined by RT-PCR and ELISA method. The necrosis area in the tumors was calculated under pathological examination. The average volume of transplanted tumor was(0.45±0.25) cm(3) in MSG-LIGHT group, (0.64±0.36) cm(3) in MSG group, and(1.21±0.79) cm(3) in NS group, and the difference was statistically significant(P<0.05). The LIGHT mRNA was 2.96±0.27, 1.23±0.47, and 0.73±0.10 respectively. The LIGHT protein was(167.89±2.31), (73.22±5.74), and (49.66±5.25) ng/L. The differences were all statistically significant among the three groups(both P<0.01). Pathological examination showed that the necrosis area was largest in MSC-LIGHT group. Transgenic UCBMSCs secret LIGHT in a paracrine manner, which has inhibition and killing effects on stomach carcinoma.

Hearing restoration in a deaf animal model with intravenous transplantation of mesenchymal stem cells derived from human umbilical cord blood

ObjectiveThis study was performed to confirm the effect of transplantation of human umbilical cord blood mesenchymal stem cells (UCB-MSCs) on hearing restoration in a sensorineural hearing loss (SNHL) animal model. Material and methodsUCB was collected from pregnant women after obtaining consent, and mesenchymal stem cells (MSCs) were extracted. We established an SNHL model and transplanted UCB-MSCs through the brachial vein of the guinea pigs. The animals were divided into 4 groups: animals with normal hearing, animals with SNHL, animals with SNHL and injected with saline, and animals with SNHL and transplanted with UCB-MSCs. Hearing tests were conducted at 1, 3, and 5weeks, and the results were compared by grading auditory brainstem response (ABR) recordings and distortion product otoacoustic emissions (DPOAEs) for each treatment. Lastly, cochlear pathological features were examined, and surface preparations and morphological changes in each animal model were compared using hematoxylin and eosin staining and light microscopy studies. ResultsIn SNHL group, decreased DPOAEs and increased ABR threshold were noted. Furthermore, in the SNHL group, ABR hearing thresholds were unconverted and were similar to those observed in deafness. The transplanted UCB-MSC group showed a significant improvement in hearing threshold (40dB) compared to that in all the SNHL group (80–90dB). Examination of the SNHL animals′ cochlear morphological features demonstrated a noticeable lack of spiral ganglion cells and also showed degenerated outer hair cells. However, the transplanted UCB-MSCs showed an increase in spiral ganglion and hair cells. ConclusionIntravenous transplantation of UCB-MSCs can enhance hearing thresholds, outer-hair cells and increase the number of spiral ganglion neurons (SGNs).

Gene therapy of gastric cancer using LIGHT-secreting human umbilical cord blood-derived mesenchymal stem cells

Mesenchymal stem cells (MSCs) have the ability to migrate into tumors and therefore are potential vehicles for the therapy of malignant diseases. In this study, we investigated the use of umbilical cord blood mesenchymal stem cells (UCB-MSCs) as carriers for a constant source of transgenic LIGHT (TNFSF14) to target tumor cells in vivo. Lentiviral vectors carrying LIGHT genes were constructed, producing viral particles with a titer of 2 × 10(8) TU/L. Fourteen days after UCB-MSCs transfected by LIGHT gene packaged lentivirus had been injected into mouse gastric cancer models, the expression levels of LIGHT mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). Then the tumors' approximate volumes were measured. The treatment with MSC-LIGHT demonstrated a strong suppressive effect on tumor growth compared to treatment with MSC and NaCl (p < 0.001). Examination of pathological sections of the tumor tissues showed that the areas of tumor necrocis in the MSC-LIGHT group were larger than those in the MSC group. Moreover, we found that MSCs with LIGHT were able to significantly induce apoptosis of tumor cells. The expression levels of LIGHT mRNA and protein were significantly higher in the UCB-MSCs with the LIGHT gene than the levels in UCB-MSCs (p < 0.001). These results suggest that UCB-MSCs carrying the LIGHT gene have the potential to be used as effective delivery vehicles in the treatment of gastric cancers.

Comparative transcriptome analysis reveals a fetal origin for mesenchymal stem cells and novel fetal surface antigens for noninvasive prenatal diagnosis

Objective Mesenchymal stem cells (MSCs) are an attractive source for providing the cells necessary for regenerating damaged tissues. Fetal MSCs (fMSCs) are known to proliferate fast and have an excellent osteogenic capacity, yet the underlying mechanisms need to be explored. A better understanding of MSCs from different anatomic origins and ages will eventually benefit cell-based therapies, as well as subsequent mechanistic studies in the field of stem cell biology. Materials and Methods We identified the molecular signatures of fetal and adult MSCs via a meta-analytic strategy and compared the enriched canonical pathways and genetic networks within each signature. Results Fetal MSCs were found to express more cell cycle genes, which is consistent with the results of wetlab functional assays. In addition, the genes involved in vasculogenesis, neurogenesis, Wnt, MAPKKK pathways, and RNA splicing were found to be enriched in fMSCs. Correlating with the overexpression of multilineage differentiation genes, fMSCs share more genes with embryonic stem cells (ESCs) and are, therefore, more primitive. Further exploration into the transcriptome similarities revealed that MSCs from umbilical cord blood (UCB) express dominant fMSC genes, but not adult genes, suggesting a fetal origin for UCB MSCs. Novel surface proteins that were dominantly expressed in fetal and UCB MSCs, but not in adult MSCs or maternal PBMCs, were also identified. Conclusion Our results systematically revealed the underlying genes and regulatory networks of two MSCs from unique origins, the resulting phenotypes, as well as the origin of UCB MSCs. The novel membrane proteins on the fetal MSC surface are promising candidate biomarkers for positively isolating fetal MSCs from maternal blood for noninvasive prenatal diagnosis.

A Microfluidic Platform for Multi-Antigen Immunofluorescence Assays

Traditional cell-based assays such as cell immunoassay that utilizes plastic (chamber slides, dishes, microtiter plates), Magnetic bead, enzyme-linked immunsorbent assays (ELISA) [1], FACS cell sorting is labor intensive, time consuming, and requires a large numbers of cells or reagents. In this report, a microfluidic device integrated with cell culture, washing, fixation, and antigen-antibody reaction is presented for high-throughput immunoassay. Using this microfluidic device, each assay can be performed on a small number of cells and nanolitre or picolitre of reagents, this is particularly beneficial for rare or expensive cell types such as stem cells, or flow sorted cell populations. The capability of the microfluidic device was demonstrated for seeding human umbilical cord blood mesenchymal stem cells (UC-MSCs) in chambers and detecting the expression of surface markers (CD34, CD44, CD45, CD73, CD105, HLA-DR) by immunofluorescence assay.

The mixed human umbilical cord blood-derived mesenchymal stem cells show higher antitumor effect against C6 cells than the single in vitro

Objective: It is difficult for the treatment of neurologic tumors with current therapies, including glioma. Despite the advances in cancer therapeutics, the outcomes in these patients remain poor and, therefore, new modalities are required. Recent findings have demonstrated that human umbilical cord blood mesenchymal stem cells (UCB-MSCs) have the potential to inhibit glioma cell growth in vitro. Caspase-3 plays a critical role as an executioner of apoptosis and the level of caspase-3 expression determines the degree of apoptosis in cancer cell lines. CD133 (+) glioma displays a strong resistance to chemotherapy. Interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) are important cytokines in the generation of antitumor immunity.Methods: UCB-MSCs were harvested by density gradient separation. Green fluorescence protein stable C6 cells were established. Cytotoxicity was detected by visual survival cell assay, and caspase-3 activity was assessed using immunohistochemistry staining and western blot. Flow cytometry was used to test CD133 positive C6 cells. The concentrations of IL-2 and IFN-gamma proteins secreted from UCB-MSCs were then quantified using enzyme-linked immunosorbent assay. The cytotoxicities of IL-2 alone, IFN-gamma alone, and combination of IL-2 and IFN-gamma were compared against malignant glioma cells.Results: We noted a significant cytotoxicity of UCB-MSCs for malignant glioma cells. In addition, the toxicity of mixed UCB-MSCs was significantly higher than that of single UCB-MSCs for C6 glioma cells. Capase-3 levels in UCB-MSCs treatment at an effector/target (E/T) ratio of (5+5):1 were higher than those at an E/T ratio of 10:1. On the contrary, there was no change in the protein expression of capase-3 at an E/T ratio of 0:1. Immunohistochemistry staining and western blot revealed that the expression of capase-3 was higher in mixed UCB-MSCs treatment, when compared with single UCB-MSCs. We identified reductions of approximately 39 and 73% in the number of CD133 positive C6 cells treated with the single (10:1) and the mixed [(5+5):1] UCB-MSCs respectively as compared to the control group (0:1) in transwell inserts. However, the mixed UCB-MSCs secreted more immune response-related proteins (IL-2 and IFN-gamma) than the single UCB-MSCs. Combination of IL-2 and IFN-gamma might prove more cytotoxic on target cells than IL-2 alone and IFN-gamma alone.Discussion: The data collected herein confirm for the first time that the mixed UCB-MSCs were shown to have more cytotoxic effects than the single UCB-MSCs through increasing the expression of caspase-3 and decreasing the expression of CD133 in C6 glioma cells. In addition, the mixed UCB-MSCs secrete more immune response-related proteins (IL-2 and IFN-gamma) than the single UCB-MSCs. Combination of IL-2 and IFN-gamma was shown to have more cytotoxic effects than IL-2 alone and IFN-gamma alone. These results demonstrate that the mixed UCB-MSCs are a potential new therapeutic agent for glioma.