Cord Blood-derived Mesenchymal Stem Cells Research Articles

Effects of hepatocyte growth factor gene-transfected mesenchymal stem cells on dimethylnitrosamine-induced liver fibrosis in rats

Nowadays, transplantation of human mesenchymal stem cells (MSCs) has emerged as a potential cellular therapy for liver cirrhosis. Hepatocyte growth factor (HGF) plays an important role in the regeneration of the liver. The objective of the study was to investigate the therapeutic effect of HGF-transfected human umbilical cord blood-derived MSCs on dimethylnitrosamine (DMN)-induced liver fibrosis in rats. HGF-transfected MSCs were transplanted into rats with DMN-induced liver fibrosis. H2O2-induced cytotoxicity, apoptosis and intracellular reactive oxygen species were reduced in HGF-transfected MSCs in HGF-transfected MSCs. Pro-apoptotic proteins, such as cleaved poly (ADP-ribose) polymerase and cleaved caspase-3, were decreased in HGF-transfected MSCs. Biochemical analysis showed that the levels of aspartate aminotransferase and alanine aminotransferase were decreased after transplantation of HGF-transfected MSCs in rat fibrosis. Trichrome staining showed that HGF-transfected MSCs reduced liver damage. Taken together, our study indicated that HGF-transfected MSCs have therapeutic effects on DMN-induced liver fibrosis in rats.

HUMAN UMBILICAL CORD BLOOD DERIVED MESENCHYMAL STEM CELLS REDUCE PHOSPHORYLATION AND AGGREGATION OF TAU IN AN ALZHEIMER’S DISEASE MODEL

In Alzheimer's Disease (AD), administration of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) mitigate hyperphosphorylation and aggregation of tau as well as improve impairment of spatial learning and memory in AD model. hUCB-MSCs were isolated and expanded from umbilical cord blood. The 5xFAD model rapidly develops severe amyloid pathology and appears cognitive impairment at 4 to 6 months-of-age. All assays were qualified or quantified by thioflavin T assay, immunofluorescence, western blot analysis, enzyme-linked immunosorbent assays and T-maze test. hUCB-MSCs have inhibitory effect on tau aggregation through in vitro aggregation assays using K18 fragment of human tau. Remarkably, hUCB-MSCs specifically decreases autophosphorylation of tyrosine 216 on the GSK-3β residue, as a kinase to promote phosphorylation of tau. Moreover, hUCB-MSCs effectively decreased phosphorylated forms of Tau at Thr181, Thr231 and Ser404, whereas effects of total Tau proteins on its stability by hUCB-MSCs were not detected. Strikingly, these phenomena mentioned above lead to results that paired helical filaments (PHF) of tau in the brain of 5XFAD mice diminished by treatment of hUCB-MSCs. Additionally, in T-maze testing, administered mice support that hUCB-MSCs have behavioral improvement of memory tasks most likely to occur in AD model. hUCB-MSCs diminish the phosphorylation and aggregation of tau through regulation of hUCB-MSCs, thus elucidating a therapeutic mechanism for AD.

Transplantation of stem cells from umbilical cord blood as therapy for type I diabetes.

In recent years, human umbilical cord blood has emerged as a rich source of stem, stromal and immune cells for cell-based therapy. Among the stem cells from umbilical cord blood, CD45+ multipotent stem cells and CD90+ mesenchymal stem cells have the potential to treat type I diabetes mellitus (T1DM), to correct autoimmune dysfunction and replenish β-cell numbers and function. In this review, we compare the general characteristics of umbilical cord blood-derived multipotent stem cells (UCB-SCs) and umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) and introduce their applications in T1DM. Although there are some differences in surface marker expression between UCB-SCs and UCB-MSCs, the two cell types display similar functions such as suppressing function of stimulated lymphocytes and imparting differentiation potential to insulin-producing cells (IPCs) in the setting of low immunogenicity, thereby providing a promising and safe approach for T1DM therapy.

Thrombin Preconditioning Boosts Biogenesis of Extracellular Vesicles from Mesenchymal Stem Cells and Enriches Their Cargo Contents via Protease-Activated Receptor-Mediated Signaling Pathways.

We investigated the role of protease-activated receptor (PAR)-mediated signaling pathways in the biogenesis of human umbilical cord blood-derived mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) and the enrichment of their cargo content after thrombin preconditioning. Immunoblot analyses showed that MSCs expressed two PAR subtypes: PAR-1 and PAR-3. Thrombin preconditioning significantly accelerated MSC-derived EV biogenesis more than five-fold and enriched their cargo contents by more than two-fold via activation of Rab5, early endosomal antigen (EEA)-1, and the extracellular signal regulated kinase (ERK)1/2 and AKT signaling pathways. Blockage of PAR-1 with the PAR-1-specific antagonist, SCH79797, significantly suppressed the activation of Rab5, EEA-1, and the ERK1/2 and AKT pathways and subsequently increased EV production and enriched EV cargo contents. Combined blockage of PAR-1 and PAR-3 further and significantly inhibited the activation of Rab5, EEA-1, and the ERK1/2 and AKT pathways, accelerated EV production, and enriched EV cargo contents. In summary, thrombin preconditioning boosted the biogenesis of MSC-derived EVs and enriched their cargo contents largely via PAR-1-mediated pathways and partly via PAR-1-independent, PAR-3-mediated activation of Rab5, EEA-1, and the ERK1/2 and AKT signaling pathways.

Traditional two-dimensional mesenchymal stem cells (MSCs) are better than spheroid MSCs on promoting retinal ganglion cells survival and axon regeneration

The loss of retinal ganglion cells (RGCs) is one of the common pathological features associated with optic nerve diseases leading to blindness. The aims of our study were to compare the neuroprotection of two forms of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) on RGCs and axon regeneration after optic nerve crush (ONC) in vivo, and to investigate the molecular mechanism. The effects of intravitreally transplanted hUCB-MSCs cultured in two-dimensional (2D-MSCs) and spheroids (3D-MSCs) were assessed by the survival of RGCs, regenerating axons, and flash visual evoked potentials (fVEP); the level of signal factors secreted by transplanted MSCs in vitreous and the marker protein levels of JAK/STAT3, PI3K/Akt/mTOR and MAPK/ERK pathways were detected using Bead-Based analysis and Western blot, respectively. We found that RGCs began to lose at day 3 after ONC, rapidly decreased at day 7, and flattened at day 14. The neuroprotection of transplanted 2D-MSCs was much stronger than that of 3D-MSCs. The transplanted 2D-MSCs could survive at least 2 weeks without differentiation and keep the characters of MSCs, which secreted multiple tropic factors and accompanied by activation of JAK/STAT3 and MAPK/ERK signaling pathways, top three most abundant factors: stem cell growth factor- β (SCGF-β), hepatocyte growth factor (HGF), and monocyte chemoattractant protein-1 (MCP-1). These results indicate that intravitreal injection of 2D-MSCs is a promising therapeutic strategy for retinal pathological diseases characterized by the loss of RGCs and open the door for the application of SCGF-β, HGF, and MCP-1 in the treatment of optic nerve diseases.

Mechanisms of the mixed umbilical cord blood-derived mesenchymal stem cells in inhibiting proliferation and inducing apoptosis of glioma cells

Objective To investigate the possible mechanism of mixed umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) in vitro to inhibit proliferation and induce apoptosis of C6 glioma cells. Methods The expression of B cell lymphoma/leukemia-2 (bcl-2) and cysteinyl aspartate-specific protease (Caspase)-3 proteins in C6 cells was detected by immunohistochemistry and Western blotting imprinting method, and C6 cell cycle was detected by flow cytometry. Results Different sources of UCB-MSCs have a tendency to increase the expression of apoptosis protein Caspase-3 in C6 cells and reduce the expression of apoptosis protein bcl-2. Different sources UCB-MSCs induce C6 cell apoptosis, E/T=0∶1, 5∶1, (2.5+ 2.5)∶1, 10∶1, (5+ 5)∶1 C6 cells acting on, The percentages of G0/G1 were (65.49±1.83)%, (64.83±2.19)%, (70.30±3.21)%, (69.03±2.19)%, (77.63±3.10)%, and the percentage of C6 cells in G0/G1 increased with the increase of the E/T ratio, showing a proportional dependence. Conclusion After phytohemagglutinin (PHA) stimulation, large doses of UCB-MSCs and mixed UCB-MSCs have inhibitory effects on proliferation of C6 cells in vitro, and this inhibition may be related to apoptosis induction Key words: Mesenchymal stem cells; Induce; Apoptosis

Interferon-γ-mediated secretion of tryptophanyl-tRNA synthetases has a role in protection of human umbilical cord blood-derived mesenchymal stem cells against experimental colitis

Mesenchymal stem cells (MSCs) are multipotent adult stem cells that present immunosuppressive effects in experimental and clinical trials targeting various rare diseases including inflammatory bowel disease (IBD). In addition, recent studies have reported tryptophanyl-tRNA synthetase (WRS) possesses uncanonical roles such as angiostatic and anti-inflammatory effects. However, little is known about the function of WRS in MSC-based therapy. In this study, we investigated if a novel factor, WRS, secreted from MSCs has a role in amelioration of IBD symptoms and determined a specific mechanism underlying MSC therapy. Experimental colitis was induced by administration of 3% DSS solution to 8-week-old mice and human umbilical cord blood-derived MSCs (hUCB-MSCs) were injected intraperitoneally. Secretion of WRS from hUCB-MSCs and direct effect of WRS on isolated CD4+ T cells was determined via in vitro experiments and hUCB-MSCs showed significant therapeutic rescue against experimental colitis. Importantly, WRS level in serum of colitis induced mice decreased and recovered by administration of MSCs. Through in vitro examination, WRS expression of hUCB-MSCs increased when cells were treated with interferon-γ (IFN-γ). WRS was evaluated and revealed to have a role in inhibiting activated T cells by inducing apoptosis. In summary, IFN-γ-mediated secretion of WRS from MSCs has a role in suppressive effect on excessive inflammation and disease progression of IBD and brings new highlights in the immunomodulatory potency of hUCB-MSCs.

PPIA, HPRT1, and YWHAZ Genes Are Suitable for Normalization of mRNA Expression in Long-Term Expanded Human Mesenchymal Stem Cells.

Long-term expansion of mesenchymal stem cells (MSCs) under defined culture conditions is necessary in human stem cell therapy. However, it alters the characteristics of MSCs. Since quantitative real time polymerase chain reaction (qRT-PCR) is widely used as one of the key analytical methods for comparative characterization, the validation of reference genes (RGs) for normalization under each experimental condition is important to achieve reliable qRT-PCR results. Therefore, the most stable RGs for long-term expanded bone marrow- and umbilical cord blood-derived MSCs (BM-MSCs and UCB-MSCs) under defined culture conditions for up to 20 passages were evaluated. The more apparent alterations in characteristics such as differentiation capacity, proliferation, senescence, and the expression of RGs were noted in BM-MSCs than UCB-MSCs during long-term expansion. The RG validation programs (GeNorm and NormFinder) suggested that PPIA, HPRT1, and YWHAZ were suitable for normalization in qRT-PCR regardless of MSC types and long-term culture expansion, and the traditional RGs (ACTB and GAPDH) were less stable in long-term expanded MSCs. In addition, the use of these RGs for normalization of OCT4 expression in long-term expanded BM-MSCs showed that a less stable RG (GAPDH) showed contrasting data compared to other RGs. Therefore, the use of RGs such as PPIA, HPRT1, and YWHAZ for normalization in qRT-PCR experiments is highly recommended for long-term expanded MSCs to generate accurate and reliable data.

Endothelin-1 Augments Therapeutic Potency of Human Mesenchymal Stem Cells via CDH2 and VEGF Signaling

In our previous study, we identified differences in the levels of CDH2 and vascular endothelial growth factor (VEGF) between effective and ineffective clones of human umbilical cord blood (hUCB) mesenchymal stem cells (MSCs), with regard to the infarcted rat myocardium. In this study, we compared gene expression profiles between the effective and ineffective clones and identified that endothelin-1 (EDN1) is enriched in the effective clone. In the mechanistic analyses, EDN1 significantly increased expression of CDH2 and VEGF through endothelin receptor A (EDNRA), which was prevented by EDNRA blocker, BQ123. To decipher how EDN1 induced gene expression of CDH2, we performed a promoter activity assay and identified GATA2 and MZF1 as inducers of CDH2. EDN1 significantly enhanced the promoter activity of the CDH2 gene, which was obliterated by the deletion or point mutation at GATA2 or MZF1 binding sequence. Next, therapeutic efficacy of EDN1-priming of hUCB-MSCs was tested in a rat myocardial infarction (MI) model. EDN1-primed MSCs were superior to naive MSCs at 8 weeks after MI in improving myocardial contractility (p < 0.05), reducing fibrosis area (p < 0.05), increasing engraftment efficiency (p < 0.05), and improving capillary density (p < 0.05). In conclusion, EDN1 induces CDH2 and VEGF expression in hUCB-MSCs, leading to the improved therapeutic efficacy in rat MI, suggesting that EDN1 is a potential priming agent for MSCs in regenerative medicine.

Exosomes Secreted by the Cocultures of Normal and Oxygen-Glucose-Deprived Stem Cells Improve Post-stroke Outcome.

Emerging stroke literature suggests that treatment of experimentally induced stroke with stem cells offered post-stroke neuroprotection via exosomes produced by these cells. Treatment with exosomes has great potential to overcome the limitations associated with cell-based therapies. However, in our preliminary studies, we noticed that the exosomes released from human umbilical cord blood-derived mesenchymal stem cells (MSCs) under standard culture conditions did not improve the post-stroke neurological outcome. Because of this apparent discrepancy, we hypothesized that exosome characteristics vary with the conditions of their production. Specifically, we suggest that the exosomes produced from the cocultures of regular and oxygen-glucose-deprived (OGD)MSCs in vitro would represent the exosomes produced from MSCs that are exposed to ischemic brain cells in vivo, and offer similar therapeutic benefits that the cell treatment would provide. We tested the efficacy of therapy with exosomes secreted from human umbilical cord blood(HUCB)-derived MSCs under in vitro hypoxic conditions on post-stroke brain damage and neurological outcome in a rat model of transient focal cerebral ischemia. We performed the TTC staining procedure as well as the neurological tests including the modified neurological severity scores(mNSS), the modified adhesive removal(sticky-tape), and the beam walking tests before ischemia and at regular intervals until 7 days reperfusion. Treatment with exosomes obtained from the cocultures of normal and OGD-induced MSCs reduced the infarct size and ipsilateral hemisphere swelling, preserved the neurological function, and facilitated the recovery of stroke-induced rats. Based on the results, we conclude that the treatment with exosomes secreted from MSCs at appropriate experimental conditions attenuates the post-stroke brain damage and improves the neurological outcome.

Mechanisms of mixed umbilical cord blood-derived mesenchymal stem cells inhibiting proliferation and inducing apoptosis of C6 glioma cells in vitro

Objective To study the mechanisms of the mixed umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) inhibiting proliferation and inducing apoptosis of C6 glioma cells in vitro. Methods Cysteinyl aspartate-specific protease (Caspase)-8, B cell lymphoma/leukemia-2 associated X protein (bax), B cell lymphoma/leukemia-2 (bcl-2) and Caspase-3 protein expression in C6 cells was analyzed by immunohistochemistry and Western blotting, respectively. Caspase-8, bax, bcl-2 and Caspase-3 mRNA levels were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Cell cycle was detected by flow cytometry. Apoptosis of C6 cells was examined by Annexin V-propidium iodide (PI) staining and terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) methods, respectively. Results The mixed UCB-MSCs had a tendency to increase the expression of Caspase-8, bax, Caspase-3 mRNA, and decrease the expression of bcl-2 mRNA. The expression of apoptosis protein and the number of apoptosis cells increased with the increase of the ratio of E/T (E: effect cells namely UCB-MSCs; T: target cells namely C6 cells), showing a proportional dependence relationship; E∶T=(5+ 5)∶1. The relative expression of Caspase-8, bax, bcl-2, and Caspase-3 mRNA in C6 cells was 0.78±0.13, 0.28±0.03, 0.70±0.08, and 0.76±0.12, respectively. When E/T=0∶1, 5∶1, (2.5+ 2.5)∶1, 10∶1, (5+ 5)∶1, the number of apoptosis C6 cells was (2.9±0.6), (3.1±0.5), (14.5±1.8), (14.3±2.2), and (22.8±4.5) cells, respectively. Conclusion After phytohemagglutinin (PHA) stimulation, the mixed UCB-MSCs could inhibit proliferation of C6 cells in vitro, which was related to induction of apoptosis of glioma cells. Key words: Glioma; Umbilical cord blood; Mesenchymal stem cells; Apoptosis; Proliferation

Intra-Articular Injection of 2 Different Dosages of Autologous and Allogeneic Bone Marrow- and Umbilical Cord-Derived Mesenchymal Stem Cells Triggers a Variable Inflammatory Response of the Fetlock Joint on 12 Sound Experimental Horses.

Osteoarthritis is a significant and costly cause of pain for both humans and horses. The horse has been identified as a suitable model for human osteoarthritis. Regenerative therapy with allogeneic mesenchymal stem cells (MSCs) is a promising treatment, but the safety of this procedure continues to be debated. The aim of this study is to evaluate the safety of intra-articular injections of allogeneic MSCs on healthy joints by comparing two different dosages and two different tissue sources, namely, bone marrow and umbilical cord blood, with a placebo treatment on the same individuals. We also assessed the influence of autologous versus allogeneic cells for bone marrow-derived MSC treatment. Twelve clinically sound horses were subjected to injections in their 4 fetlock joints. Each of the three fetlocks was administered a different MSC type, and the remaining fetlock was injected with phosphate-buffered saline as a control. Six horses received 10 million cells per joint, and the 6 other horses received 20 million cells per joint. Clinical and ultrasound monitoring revealed that allogeneic bone marrow-derived MSCs induced significantly more synovial effusion compared to umbilical cord blood-derived MSCs but no significant difference was noted within the synovial fluid parameters. The administration of 10 million cells in horses triggered significantly more inflammatory signs than the administration of 20 million cells. Mesenchymal stem cell injections induced mild to moderate local inflammatory signs compared to the placebo, with individual variability in the sensitivity to the same line of MSCs. Understanding the behavior of stem cells when injected alone is a step towards the safer use of new strategies in stem cell therapy, where the use of either MSC secretome or MSCs combined with biomaterials could enhance their viability and metabolic activity.

TGF-beta in the UCB-MSCs inhibit TNF-alpha in human mast cells and atopic dermatitis

Abstract It is well known that umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) have been effective therapy in autoimmune inflammatory disease such as atopic dermatitis (AD), rheumatoid arthritis (RA) and psoriasis. Especially, UCB-MSC has a prominent therapeutic effect on AD. However, there is not much research on the mechanism of alleviation of AD by UCB-MSCs. In this study, we investigated the mechanism of AD therapy by UCB-MSCs, using human mast cells as the major pathogenic cells in AD. LAD2 cells were stimulated with IL- 33(30ng/ml) and Ionomycin (1uM) under IgE-XL condition and they secreted a significant amount of TNF-a. TNFa was significantly inhibited by co-culture with UCB-MSCs. UCB-MSCs secreted TGF-beta and PGE2, which play a crucial role in immune regulation. TNF-a of LAD2 was increased when co-cultured with UCB-MSCs after TGF-b siRNA transfection. Nevertheless, inhibition of PGE2 using indomethacin, NS398 and celecoxib did not restore secretion of TNF-a. At the protein level, we examined the ERK, JNK and AKT signaling pathways involved in TNF-α, and only the ERK signaling pathway was associated with TNF-α inhibition. Finally, we found that TNFa is regulated by TGFb induced by UCB-MSC in a DF-induced mouse model. Taken together, our findings suggest that TGF-b may play an important role in regulating mast cells in AD rather than PGE2 in UCB-MSCs.

Conditioned Media of Human Umbilical Cord Blood Mesenchymal Stem Cell Inhibits Ultraviolet B-induced Apoptosis in Fibroblasts

BACKGROUND: Ultraviolet (UV)B irradiated-skin cells may respond either by activating protective or apoptotic mechanisms. Several treatments has been reported to prevent apoptosis of the skin cell. To date, despite of the cell, secretome of mesenchymal stem cell (MSC) has been progressively explored for various treatment. Current research was conducted to find out the possible effect of MSC's secretome to protect normal cells from the UVB irradiation.METHODS: For preparation of the conditioned media human umbilical cord blood mesenchymal stem cell (CM-hUCB-MSC), hUCB was collected and separated to collect mononuclear cell (MNC). MNC was cultured in MSC medium until passage 4, then checked for MSC biomarkers. Conditioned media was then produced from the cultured MSC. For induction of apoptosis, NIH3T3 cells were pretreated with/without CM-hUCB-MSC, prior to the UVB irradiation for 5-60 minutes. After 24 hours, apoptosis evaluation was conducted with Sub-G1 assay with hypotonic fluorochrome solution using flow cytometer.RESULTS: The average of apoptotic cells irradiated with UVB for 30 minutes was 94.2%. The 30-minutes-UVB irradiation significantly induced apoptosis in NIH3T3 cells (p=0.000). The averages of apoptotic cells by pretreatment of 0, 5, 10 and 20% CM-hUCB-MSC prior to UVB irradiation were 94.2, 46.8, 31.8 and 31.5 %, respectively. The pretreatment of 5, 10 and 20% CM-hUCB-MSC prior to UVB irradiation could significantly decrease the percentage of apoptosis caused by UVB (p=0.001).CONCLUSION: Taken together, CM-hUCB-MSC inhibited UVB-induced apoptosis in NIH3T3 cells significantly, suggesting that CM-hUCB-MSC might be a potential anti-UVB-induced apoptosis factor. Hence, further research should be explored to disclose its specific intracellular mechanism.KEYWORDS: UVB, stem cell, conditioned media, fibroblast, apoptosis, aging, secretome, NIH3T3 cell

Clinical outcomes of human umbilical cord blood derived mesenchymal stem cells application in knee osteoarthritis patients

Purpose: There are many animal studies on the articular cartilage regeneration using stem cells, but studies on stem cell treatment on human are rare. For patients with knee osteoarthritis with full thickness cartilage defect (> 2.0 cm2), we have been using human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) to regenerate articular cartilage since 2012. The purpose of this study is to analyze the effect of hUCB-MSCs based cartilage regeneration surgery on the preoperative and postoperative changes of symptoms, function and arthroscopic findings. Methods: After standard arthroscopic examination, the cartilage defect site was exposed through small arthrotomy or arthroscopy. Then, multiple drill holes (2-5mm in diameter and 5mm depth) were made approximately 2-3 mm apart at the cartilage defect site, and hUCB-MSCs mixture was implanted in the drill holes of the lesion. Patients with varus or valgus deformity greater than 5 degrees were treated simultaneously with osteotomy. Among 253 patients (276 knees) who underwent hUCB-MSCs surgery from February 2012 to October 2017, we reviewed 64 patients (76 knees) who underwent second-look arthroscopy after postoperatively 1 year. Cartilage regeneration was graded according to Oswestry Arthroscopy Score (OAS). Functional outcomes using subjective International Knee Documentation Committee (IKDC) score, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score, and visual analog scale (VAS) were evaluated at preoperative and postoperative 1 year. Results: Totally 76 knees (64 patients; mean age, 59.8 ± 8.7 years; age range, 31 to 79 years) underwent second-look arthroscopic surgery at postoperative 1 year. The average preoperative cartilage defect is 4.4 ± 1.1 cm2 (range, 2.0 to 5.9 cm2). IKDC score improved from 22.9 points preoperatively to 69.3 points at postoperative 1 year. WOMAC score (mean 52.3 points preoperatively to 13.4 points at postoperative 1 year) and VAS score (mean 7.7 points preoperatively to 1.9 points at postoperative 1 year) all significantly improved at postoperative 1 year. The mean OAS were significantly improved from 5.6 ± 1.8 preoperatively to 2.1 ± 2.4 at postoperative 1 year. There was no complication due to hUCB-MSCs application itself, but in 2 kness, there was poor cartilage regeneration, pain persisted, so converted to TKA. Conclusions: Although there is no histological study and lifespan of the regenerated cartilage, the cartilage regeneration using hUCB-MSCs showed significant pain and function improvement, and excellent arthroscopic findings of cartilage regeneration. These results suggest that hUCB-MSCs application surgery is good treatment option for patients with severe knee osteoarthritis of relatively younger age to perform TKA surgery.

Improved expansion of equine cord blood derived mesenchymal stromal cells by using microcarriers in stirred suspension bioreactors

Equine mesenchymal stromal cells (MSCs) are increasingly investigated for their clinical therapeutic utility. Such cell-based treatments can require cell numbers in the millions or billions, with conventional expansion methods using static T-flasks typically inefficient in achieving these cell numbers. Equine cord blood-derived MSCs (eCB-MSCs), are promising cell candidates owing to their capacity for chondrogenic differentiation and immunomodulation. Expansion of eCB-MSCs in stirred suspension bioreactors with microcarriers as an attachment surface has the potential to generate clinically relevant numbers of cells while decreasing cost, time and labour requirements and increasing reproducibility and yield when compared to static expansion. As eCB-MSCs have not yet been expanded in stirred suspension bioreactors, a robust protocol was required to expand these cells using this method. This study outlines the development of an expansion bioprocess, detailing the inoculation phase, expansion phase, and harvesting phase, followed by phenotypic and trilineage differentiation characterization of two eCB-MSC donors. The process achieved maximum cell densities up to 75,000 cells/cm2 corresponding to 40 million cells in a 100 mL bioreactor, with a harvesting efficiency of up to 80%, corresponding to a yield of 32 million cells from a 100 mL bioreactor. When compared to cells grown in static T-flasks, bioreactor-expanded eCB-MSC cultures did not change in surface marker expression or trilineage differentiation capacity. This indicates that the bioreactor expansion process yields large quantities of eCB-MSCs with similar characteristics to conventionally grown eCB-MSCs.

Photobiomodulation Enhances the Angiogenic Effect of Mesenchymal Stem Cells to Mitigate Radiation-Induced Enteropathy.

Radiation-induced enteropathy remains a major complication after accidental or therapeutic exposure to ionizing radiation. Recent evidence suggests that intestinal microvascular damage significantly affects the development of radiation enteropathy. Mesenchymal stem cell (MSC) therapy is a promising tool to regenerate various tissues, including skin and intestine. Further, photobiomodulation (PBM), or low-level light therapy, can accelerate wound healing, especially by stimulating angiogenesis, and stem cells are particularly susceptible to PBM. Here, we explored the effect of PBM on the therapeutic potential of MSCs for the management of radiation enteropathy. In vitro, using human umbilical cord blood-derived MSCs, PBM increased proliferation and self-renewal. Intriguingly, the conditioned medium from MSCs treated with PBM attenuated irradiation-induced apoptosis and impaired tube formation in vascular endothelial cells, and these protective effects were associated with the upregulation of several angiogenic factors. In a mouse model of radiation-induced enteropathy, treatment with PBM-preconditioned MSCs alleviated mucosal destruction, improved crypt cell proliferation and epithelial barrier functions, and significantly attenuated the loss of microvascular endothelial cells in the irradiated intestinal mucosa. This treatment also significantly increased angiogenesis in the lamina propria. Together, we suggest that PBM enhances the angiogenic potential of MSCs, leading to improved therapeutic efficacy for the treatment of radiation-induced enteropathy.

WITHDRAWN: Highlights – Improved osteogenic differentiation of umbilical cord blood MSCs using custom made perfusion bioreactor

WITHDRAWN: Highlights – Improved osteogenic differentiation of umbilical cord blood MSCs using custom made perfusion bioreactor

CRISPR/Cas9 Edited sRAGE-MSCs Protect Neuronal Death in Parkinson’s Disease Model.

Background and ObjectivesParkinson’s disease (PD) is a fatal and progressive degenerative disease of the nervous system. Until recently, its promising treatment and underlying mechanisms for neuronal death are poorly understood. This study was investigated to identify the molecular mechanism of neuronal death in the substantia nigra and corpus striatum of PD.MethodsThe soluble RAGE (sRAGE) secreting Umbilical Cord Blood—derived Mesenchymal Stem Cell (UCB-MSC) was generated by gene editing method using clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9 (CRISPR/Cas9). These cells were transplanted into Corpus Striatum of rotenone-induced PD animal models then behavioral test, morphological analysis, and immunohistochemical experiments were performed to determine the neuronal cell death and recovery of movement.ResultsThe neuronal cell death in Corpus Striatum and Substantia Nigra was dramatically reduced and the movement was improved after sRAGE secreting UCB-MSC treatment in PD mice by inhibition of RAGE in neuronal cells.ConclusionsWe suggest that sRAGE secreting UCB-MSC based therapeutic approach could be a potential treatment strategy for neurodegenerative disease including PD.

Umbilical cord blood mesenchymal stem cells application in hematopoietic stem cells expansion on nanofiber three-dimensional scaffold.

Umbilical cord blood (UCB) hematopoietic stem cells (HSCs) transplantation (HSCTs) is considered as a therapeutic strategy for malignant and nonmalignant hematologic disorders. Nevertheless, the low number of HSCs obtained from each unit of UCB can be a major challenge for using these cells in adults. In addition, UCB is a rich source of mesenchymal stem cells (MSCs) creating hopes for nonaggressive and painless treatment in tissue engineering compared with bone marrow MSCs. This study was designed to evaluate the effects of UCB-MSCs application in UCB-HSCs expansion on the nanoscaffold that mimics the cell's natural niche. To achieve this goal, after flow cytometry confirmation of isolated HSCs from UCB, they were expanded on three-dimensional (3D) poly-l-lactic acid (PLLA) scaffolds fabricated by electrospinning and two-dimensional (2D)-culture systems, such as (1) HSCs-MSCs culturing on the scaffold, (2) HSCs culturing on the scaffold, (3) HSCs-MSCs culturing on 2D, and (4) HSCs culturing on 2D. After 7 days, real-time polymerase chain reaction (PCR) was performed to evaluate the CXCR4 gene expression in the mentioned groups. Moreover, for the next validation, the number of total HSCs, 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide assay, scanning electron microscopy imaging, and colony-forming unit assay were evaluated as well. The results of the study indicated that UCB-MSCs interaction with HSCs in 3D-culture systems led to the highest expansion of UCB-HSCs on day 7. Flow cytometry results showed the highest purity of HSCs cocultured with MSCs. Real-time PCR showed a significant increase in gene expression of CXCR4 in the mentioned group. The highest viability and clonogenicity were detected in the mentioned group too. Considered together, our results suggest that UCB-HSCs and MSCs coculturing on PLLA scaffold could provide a proper microenvironment that efficiently promotes UCB-HSCs expansion and UCB-MSCs can also be considered as a promising candidate for UCB-HSCTs.