Umbilical Cord Blood-derived Mesenchymal Stem Cells Research Articles

THE INFLUENCE OF MESENCHYMAL STROMAL CELLS OF DIFFERENT GENESIS ON ENERGY METABOLISM IN THE RAT SOMATOSENSORY CORTEX DURING ISCHEMIA-REPERFUSION

Introduction. About 60-80% of cerebral circulation disorders are due to ischemia, that leads to high mortality, disability in working age and economic burden on society in developed countries of the world. Objective: To investigate the effect of ischemia-reperfusion on parameters of carbohydrate and energy exchanges in the rat somatosensory cortex and to determine the cerebroprotective action of mesenchymal stromal cells (MSCs) of various genesis, MSC lysate and the reference drug Citicoline. Methods. The study was performed on 200 male Wistar rats, divided into 9 groups. Group 1 included intact animals; group 2 – sham-operated rats; group 3 – control pathology (rats were subjected to ischemia-reperfusion by occlusion of the internal carotid arteries and injected with a 0.9% saline solution (2 ml/kg) into the femoral vein); group 4 – immediately after ischemia-reperfusion rats were transplanted with 106 MSC cells/animal derived from Wharton’s jelly of the human umbilical cord; group 5 – 106 cells/animal of rat embryonic fibroblasts; group 6 – 106 cells/animal derived from human adipose MSCs; group 7 – 106 cells/animal derived from rat adipose MSCs; group 8 – 0.2 ml/animal of lysate derived from Wharton’s jelly MSCs; group 9 – the reference drug Citicoline (250 mg/kg). Parameters of carbohydrate and energy metabolism in rat somatosensory cortex under the conditions of cerebral IR and on the background of correction studied on the 7th and 14th day. Results. It was found that after cerebral ischemia-reperfusion in the rat somatosensory cortex in research periods an increase in the levels of glucose, lactate, lactate/pyruvate ratio and a decrease in the content of pyruvate and succinate dehydrogenase were revealed. In the groups of experimental animals with intravenous transplantation of MSCs of various origins, MSC lysate and Citicoline, the perturbation in glucose metabolism was reduced, lactate content was decreased and the levels of pyruvate and succinate dehydrogenase were significantly increased (p<0.05), compared to rats with control pathology. A more pronounced positive effect in the recovery of disturbed energy processes and elimination of metabolic acidosis was observed with the use of human Wharton’s jelly-derived MSCs. Conclusions. The study demonstrated that experimental 20-minute model ischemia-reperfusion in rats caused a violation of the carbohydrate and energy balance in the somatosensory cortex. Human umbilical cord blood-derived MSCs contributed to the recovery of disturbed energy processes in the somatosensory cortex and eliminated metabolic acidosis at the level of Citicoline or better than it, which is one of the links of the mechanism of the cerebral protective action of MSCs. Embryonic rat fibroblasts were slightly inferior in efficiency to Citicoline and human umbilical cord Wharton’s jelly MSCs, which indicates a higher cerebroprotective activity of xenotransplantation.

Tacrolimus Improves Therapeutic Efficacy of Umbilical Cord Blood-Derived Mesenchymal Stem Cells in Diabetic Retinopathy by Suppressing DRP1-Mediated Mitochondrial Fission.

Diabetic retinopathy (DR) is a leading cause of blindness in diabetic patients. Umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) are emerging as a promising new drug for degenerative disease associated with diabetes. Recent studies have shown that high glucose-increased excessive calcium levels are a major risk factor for mitochondrial reactive oxygen species (mtROS) accumulation and apoptosis. This study aimed to investigate the role of high glucose-induced NFATC1 signaling in mitochondrial oxidative stress-stimulated apoptosis and the effect of tacrolimus on the therapeutic efficacy of subconjunctival transplantation of UCB-MSCs in a DR rat model. High glucose increased mtROS and cleaved caspase-9 expression in UCB-MSCs. High glucose conditions increased O-GlcNAcylated protein expression and nuclear translocation of NFATC1. Tacrolimus pretreatment recovered high glucose-induced mtROS levels and apoptosis. In the DR rat model, subconjunctival transplantation of tacrolimus-pretreated MSCs improved retinal vessel formation, retinal function, and uveitis. In high glucose conditions, tacrolimus pretreatment reduced protein and mRNA expression levels of DRP1 and inhibited mitochondrial fission. In conclusion, we demonstrated that high glucose-induced O-GlcNAcylation activates NFATC1 signaling, which is important for DRP1-mediated mitochondrial fission and mitochondrial apoptosis. Finally, we proposed NFATC1 suppression by tacrolimus as a promising therapeutic strategy to improve the therapeutic efficacy of UCB-MSC transplantation for DR treatment.

A Comparative Histological Study on the Role of Umbilical Cord Mesenchymal Stem Cells versus Their Conditioned Medium on the Pancreatic Beta Cells in Experimentally Induced Diabetes Mellitus in Albino Rat

Abstract Background Mesenchymal stem cells (MSCs) represent a promising population for supporting new clinical concepts in cellular therapy. Umbilical cord blood (UCB) has turned out to be an excellent alternative source of MSCs. This study had compared the role of human umbilical cord blood derived mesenchymal stem cells (UCB-MSCs) versus their conditioned medium (CM) alone on pancreatic beta cells in a rat model of Streptozotocin (STZ)-induced type I diabetes Mellitus. Material and Methods Forty adult male albino rats were divided randomly into 4 groups; Group I (control), Group II (STZ), rats received a single dose of intraperitoneal injection of 35 mg/kg of STZ dissolved in 1 ml sodium citrate buffer. Group III (STZ + UCB-MSCs) after rats had been confirmed to be diabetic they received single dose of 1 × 106 cells /ml of UCB-MSCs into tail vein, meanwhile Group IV (STZ+ CM) rats received 0.5 ml of CM that was injected intramuscularly once per week for 4 weeks. After 4 weeks the Pancreas from all groups were dissected and stained by Hematoxylin & eosin stain and by immunohistochemical method using anti-insulin antibody to detect insulin granules in the Beta cells of the islets. Mean area percent of insulin positive reactions were measured and statistically analyzed. Results H&E-stained section of group II (STZ) showed the nuclei of the islet cells small condensed and darkly stained while, the blood capillaries were congested in endocrine pancreas.Meanwhile, Group III (STZ + UCB-MSCs) restored nearly normal histological features. Sections of Group IV (STZ+CM) showed few cells of the islets sections of showed vacuolations and small darkly stained condensed nuclei. Studying the groups immunohistochemically, revealed decrease in insulin positive reactions in Group II (STZ) while an increase in insulin positive reactions noticed in group III (STZ +UCB-MSC) and group IV (STZ+CM) when compared to group II (STZ) with more increase in group III (STZ+UCB-MSC) than group IV (STZ+CM). Conclusion It can be considered that intravenous UCB-MSCs could be more effective than CM in treatment of STZ-induced type I diabetes in the pancreas.

Clinical and Magnetic Resonance Imaging Outcomes After Human Cord Blood–Derived Mesenchymal Stem Cell Implantation for Chondral Defects of the Knee

There is a paucity of literature reporting clinical and magnetic resonance imaging (MRI) outcomes after allogeneic umbilical cord blood-derived mesenchymal stem cell (UCB-MSC) implantation for chondral defects of the knee. To report clinical and MRI outcomes after UCB-MSC implantation for chondral lesions of the knee. Case series; Level of evidence, 4. Inclusion criteria were patients aged between 40 and 70 years with focal chondral lesions of grade 3 or 4 on the medial femoral condyle, defect sizes >4 cm2, and intact ligaments. Exclusion criteria were patients who required realignment osteotomy or who had a meniscal deficiency, ligamentous instability, or a concomitant full-thickness chondral defect in the lateral or patellofemoral compartment. A mixture of human UCB-MSCs and sodium hyaluronate was implanted into the chondral defect through mini-arthrotomy. MRI at 1-year follow-up was performed to evaluate repaired cartilage hypertrophy. Repaired cartilage thickness was measured, and hypertrophy was classified as grade 1 (<125%), grade 2 (<150%), or grade 3 (<200%). Patient-reported outcomes (PROs; International Knee Documentation Committee, visual analog scale for pain, and Western Ontario and McMaster Universities Osteoarthritis Index) were evaluated preoperatively and at 1, 2, and 3 years postoperatively. Repaired cartilage hypertrophy was evaluated for a correlation with PRO scores. Enrolled were 85 patients with a mean age of 56.8 ± 6.1 years and a mean chondral defect size of 6.7 ± 2.0 cm2. At follow-up, a significant improvement in all PRO scores was seen compared with preoperatively (P < .001 for all). MRI at 1-year follow-up demonstrated that 28 patients had grade 1 repaired cartilage hypertrophy, 41 patients had grade 2, and 16 patients had grade 3. MRI performed in 11 patients at 2 years after surgery indicated no difference in repaired cartilage hypertrophy between the 1- and 2-year time points. The grade of repaired cartilage hypertrophy did not correlate with PRO scores. Clinical outcomes improved significantly at short-term follow-up after UCB-MSC implantation. Although all patients showed repaired cartilage hypertrophy, it did not correlate with clinical outcomes.

Bone Marrow Aspirate Concentrate versus Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells for Combined Cartilage Regeneration Procedure in Patients Undergoing High Tibial Osteotomy: A Systematic Review and Meta-Analysis.

Background and objectives: Cartilage regeneration using mesenchymal stem cells (MSCs) has been attempted to improve articular cartilage regeneration in varus knee osteoarthritis (OA) patients undergoing high tibial osteotomy (HTO). Bone marrow aspirate concentrate (BMAC) and human umbilical cord blood-derived MSCs (hUCB-MSCs) have been reported to be effective. However, whether BMAC is superior to hUCB-MSCs remains unclear. This systematic review and meta-analysis aimed to determine the clinical efficacy of cartilage repair procedures with BMAC or hUCB-MSCs in patients undergoing HTO. Materials and Methods: A systematic search was conducted using three global databases, PubMed, EMBASE, and the Cochrane Library, for studies in which the clinical outcomes after BMAC or hUCB-MSCs were used in patients undergoing HTO for varus knee OA. Data extraction, quality control, and meta-analysis were performed. To compare the clinical efficacy of BMAC and hUCB-MSCs, reported clinical outcome assessments and second-look arthroscopic findings were analyzed using standardized mean differences (SMDs) with 95% confidence intervals (CIs). Results: The present review included seven studies of 499 patients who received either BMAC (BMAC group, n = 169) or hUCB-MSCs (hUCB-MSC group, n = 330). Improved clinical outcomes were found in both BMAC and hUCB-MSC groups; however, a significant difference was not observed between procedures (International Knee Documentation Committee score; p = 0.91, Western Ontario and McMaster Universities OA Index; p = 0.05, Knee Society Score (KSS) Pain; p = 0.85, KSS Function; p = 0.37). On second-look arthroscopy, the hUCB-MSC group showed better International Cartilage Repair Society Cartilage Repair Assessment grade compared with the BMAC group (p < 0.001). Conclusions: Both BMAC and hUCB-MSCs with HTO improved clinical outcomes in varus knee OA patients, and there was no difference in clinical outcomes between them. However, hUCB-MSCs were more effective in articular cartilage regeneration than BMAC augmentation.

Allogeneic Umbilical Cord-Blood-Derived Mesenchymal Stem Cells and Hyaluronate Composite Combined with High Tibial Osteotomy for Medial Knee Osteoarthritis with Full-Thickness Cartilage Defects.

Background and Objectives: Although the effects of cartilage repair in patients who are undergoing high tibial osteotomy (HTO) remains controversial, cartilage repair may be required for the full-thickness cartilage defect because of a concern of lower clinical outcome. The purpose of this study was to investigate clinical outcome and cartilage repair following implantation of allogeneic umbilical cord-blood-derived MSCs (UCB-MSCs)-hyaluronate composite in patients who received HTO for medial knee osteoarthritis (OA) with full-thickness cartilage defect. Materials and Methods: Inclusion criteria were patients with a medial knee OA, a full-thickness cartilage defect (International Cartilage Repair Society (ICRS) grade IV) ≥ 3 cm2 of the medial femoral condyle, and a varus deformity ≥ 5°. The full-thickness cartilage defect was treated with implantation of an allogeneic UCB-MSCs-hyaluronate composite following medial open-wedge HTO. Visual analogue scale for pain and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score were assessed at each follow-up. Cartilage repair was assessed by the ICRS cartilage repair assessment system at second-look arthroscopy when the plate was removed. Results: Twelve patients (mean age 56.1 years; mean defect size: 4.5 cm2) were included, and 10 patients underwent second-look arthroscopy during plate removal after a minimum of 1 year after the HTO. At the final follow-up of mean 2.9 years (range; 1-6 years), all clinical outcomes had improved. At second-look arthroscopy, repaired tissue was observed in all cases. One case (10%) showed grade I, seven (70%) cases showed grade II, and two (20%) cases showed grade III according to ICRS cartilage repair assessment system, which meant that 80% showed an overall repair assessment of "normal" or "nearly normal". Conclusion: Allogeneic UCB-MSCs-HA composite implantation combined with HTO resulted in favorable clinical outcome and cartilage repair in all cases. These findings suggest that UCB-MSCs-HA composite implantation combined with HTO would be a good therapeutic option for patients with knee OA and full-thickness cartilage defects.

The efficiency of human umbilical cord mesenchymal stem cells as a salvage treatment for steroid-refractory acute graft-versus-host disease.

Acute graft-versus-host disease (aGVHD) is a life-threatening complication after hematopoietic stem cell transplantation (HSCT) and is primarily treated with steroids. However, there is no standard treatment for steroid-refractory acute graft-versus-host disease (SR-aGVHD). Although mesenchymal stem cells (MSCs) have proven effective for SR-aGVHD, few reports have focused on human umbilical cord blood-derived MSCs (hUCB-MSCs). Here, we report on the efficiency of hUCB-MSCs as the salvage therapy for SR-aGVHD in 54 patients. The overall response rate (ORR) reached 59.3% (32/54) 28days later. Twenty-four patients achieved complete remission (CR), and 8 achieved partial remission (PR). The median follow-up time after the initiation of hUCB-MSC treatment was 19.3 (0.6-59.0) months. The probability of overall survival (OS) and progression-free survival (PFS) was 60.9% (47.4-74.4%, 95% CI) and 58.8% (45.3-72.3%, 95% CI), respectively, while that of GVHD/relapse-free survival (GRFS) was only 30.8% (17.86-43.74%, 95% CI). Multivariate analysis revealed that response on Day 28 was an independent favorable prognostic factor (OS, P < 0.001; PFS, P < 0.001; GRFS, P = 0.001), but an age of ≥ 18years suggested an unfavorable long-term prognosis (OS, P < 0.001; PFS, P < 0.001; GRFS, P = 0.003). In addition, liver involvement was adversely associated with PFS (P = 0.021) and GRFS (P = 0.009). An infused MNC ≥ 8.66 × 108/kg was also detrimental to GRFS (P = 0.031). Collectively, our results support hUCB-MSCs as an effective treatment for SR-aGVHD.

Comparison of the therapeutic effects of human umbilical cord blood-derived mesenchymal stem cells and adipose-derived stem cells on erectile dysfunction in a rat model of bilateral cavernous nerve injury.

Background: Cavernous nerve injury (CNI) is the leading cause of erectile dysfunction (ED) after radical prostatectomy and pelvic fracture. Transplantation of human adipose-derived stem cells (ASCs) has been widely used to restore erectile function in CNI-ED rats and patients. Umbilical cord blood-derived MSCs (CBMSCs) are similarly low immunogenic but much primitive compared to ASCs and more promising in large-scale commercial applications due to the extensive establishment of cord blood banks. However, whether CBMSCs and ASCs have differential therapeutic efficacy on CNI-ED and the underlying mechanisms are still not clear. Materials and methods: A bilateral cavernous nerve injury (BCNI) rat model was established by crushing the bilateral cavernous nerves. After crushing, ASCs and CBMSCs were intracavernously injected immediately. Erectile function, Masson staining, and immunofluorescence analyses of penile tissues were assessed at 4 and 12 weeks. PKH-26-labeled ASCs or CBMSCs were intracavernously injected to determine the presence and differentiation of ASCs or CBMSCs in the penis 3 days after injection. In vitro experiments including intracellular ROS detection, mitochondrial membrane potential assay, EdU cell proliferation staining, cell apoptosis assay, and protein chip assay were conducted to explore the underlying mechanism of CBMSC treatment compared with ASC treatment. Results: CBMSC injection significantly restored erectile function, rescued the loss of cavernous corporal smooth muscles, and increased the ratio of smooth muscle to collagen. PKH-26-labeled CBMSCs or ASCs did not colocalize with endothelial cells or smooth muscle cells in the corpus cavernosum. Moreover, the conditioned medium (CM) of CBMSCs could significantly inhibit the oxidative stress and elevate the mitochondria membrane potential and proliferation of Schwann cells. Better therapeutic effects were observed in the CBMSC group than the ASC group both in vivo and in vitro. In addition, the content of neurotrophic factors and matrix metalloproteinases in CBMSC-CM, especially NT4, VEGF, MMP1, and MMP3 was significantly higher than that of ASC-CM. Conclusion: Intracavernous injection of CBMSCs exhibited a better erectile function restoration than that of ASCs in CNI-ED rats owing to richer secretory factors, which can promote nerve regeneration and reduce extracellular matrix deposition. CBMSC transplantation would be a promising therapeutic strategy for CNI-ED regeneration in the future.

Silencing SIRT5 induces the senescence of UCB-MSCs exposed to TNF-α by reduction of fatty acid β-oxidation and anti-oxidation

Tumor necrosis factor-α (TNF-α) is an inflammatory cytokine involved in cell survival, apoptosis, and homeostasis. However, the regulatory effect of TNF-α on mesenchymal stem cell (MSC) redox regulation remains unknown. The process of delaying the senescence of MSCs and maintaining antioxidation mechanism is important in transplantation therapy to treat inflammatory diseases that result from restricted immunomodulatory effects of senescent MSCs. Thus, we examined the role of TNF-α-mediated signaling and its regulatory mechanisms on the senescence of umbilical cord blood-derived MSCs (UCB-MSCs) and identified its therapeutic efficacy in a collagen-induced arthritis (CIA) mouse model. We found that TNF-α increased fatty acid synthesis and lipid droplet (LD) formation through NF-κB/SREBP1-mediated FASN, SCD1, and DGAT2 expression, which protects UCB-MSCs from oxidative stress against accumulated toxic lipids. Additionally, DGAT2-mediated LD formation was regulated by TNF-α-activated TNF receptor (TNFR)1 signaling. We also found that storage of unsaturated FAs in LDs is regulated by SIRT5-dependent β-oxidation of FAs, which reduces mitochondrial ROS (mtROS) accumulation. Particularly, mtROS homeostasis was maintained by superoxide dismutase 2 (SOD2) upregulation through TNFR2-mediated SIRT5/Nrf2 signaling. In a CIA mouse model, UCB-MSCs transfected with SIRT5 siRNA exhibited reduced therapeutic effects compared with UCB-MSCs transfected with NT siRNA. Overall, the results indicated that SIRT5 plays a central role in protecting TNF-α-induced UCB-MSC senescence through FA β-oxidation and SOD2-mediated antioxidation.

Human umbilical cord blood-derived MSCs trans-differentiate into endometrial cells and regulate Th17/Treg balance through NF-κB signaling in rabbit intrauterine adhesions endometrium

PurposeThe fundamental cause of intrauterine adhesions (IUAs) is the destruction and reduction in stem cells in endometrial basal layer, resulting in endometrial reconstruction very difficult. The purpose of this study was to investigate the effects and underlying mechanism of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) on the endometrial reconstruction after transplantation.MethodshUCB-MSCs were isolated and identified by flow cytometry, osteogenic, adipogenic and chondrogenic differentiation assays. The rabbit IUA models were established and set five groups (control, 14/28th day after surgery, estrogen and hUCB-MSCs treatment). The number of endometrial glands and the fibrosis rate were evaluated using HE and Masson staining, respectively. Endometrial proliferation, angiogenesis and inflammation were evaluated by immunohistochemical staining of ER, Ki-67and TGF-β1, respectively. Single-cell RNA sequencing (scRNA-seq) was applied to explore the cell differentiation trajectory after hUCB-MSCs transplanted into IUA endometrium. Finally, molecular mechanism of hUCB-MSCs repairing damaged endometrium was investigated by RNA sequencing, qRT-PCR and Western blot assays.ResultsAfter transplantation of the hUCB-MSCs, the increase in endometrial gland number, estrogen receptor (ER) and Ki-67 expression, and the decrease in fibrosis rate and TGF-β expression (P < 0.05), suggested the endometrial repair, angiogenesis and inflammatory suppression. The therapeutic effect of hUCB-MSCs was significantly improved compared with 28th day after surgery and estrogen group. ScRNA-seq demonstrated that the transplanted hUCB-MSCs can trans-differentiate into endometrial cells: epithelial, fibroblast and macrophage. RNA sequencing of six IUA samples combined with qRT-PCR and Western blot assays further revealed that hUCB-MSCs may regulate Th17/Treg balance through NF-κB signaling, thus inhibiting the immune response of damaged endometrium.ConclusionsOur study demonstrated that hUCB-MSCs can repair damaged endometrium through trans-differentiation, immunomodulatory capacities and NF-κB signaling, suggesting the treatment value of hUCB-MSCs in IUA.

Intratracheal Transplantation of Mesenchymal Stem Cells Attenuates Hyperoxia-Induced Microbial Dysbiosis in the Lungs, Brain, and Gut in Newborn Rats.

We attempted to determine whether intratracheal (IT) transplantation of mesenchymal stem cells (MSCs) could simultaneously attenuate hyperoxia-induced lung injuries and microbial dysbiosis of the lungs, brain, and gut in newborn rats. Newborn rats were exposed to hyperoxia (90% oxygen) for 14 days. Human umbilical cord blood-derived MSCs (5 × 105) were transplanted via the IT route on postnatal day (P) five. At P14, the lungs were harvested for histological, biochemical, and microbiome analyses. Bacterial 16S ribosomal RNA genes from the lungs, brain, and large intestine were amplified, pyrosequenced, and analyzed. IT transplantation of MSCs simultaneously attenuated hyperoxia-induced lung inflammation and the ensuing injuries, as well as the dysbiosis of the lungs, brain, and gut. In correlation analyses, lung interleukin-6 (IL-6) levels were significantly positively correlated with the abundance of Proteobacteria in the lungs, brain, and gut, and it was significantly inversely correlated with the abundance of Firmicutes in the gut and lungs and that of Bacteroidetes in the lungs. In conclusion, microbial dysbiosis in the lungs, brain, and gut does not cause but is caused by hyperoxic lung inflammation and ensuing injuries, and IT transplantation of MSCs attenuates dysbiosis in the lungs, brain, and gut, primarily by their anti-oxidative and anti-inflammatory effects.

HLA-A2 Promotes the Therapeutic Effect of Umbilical Cord Blood-Derived Mesenchymal Stem Cells in Hyperoxic Lung Injury.

Mesenchymal stem cells (MSCs) are one of the most extensively studied stem cell types owing to their capacity for differentiation into multiple lineages as well as their ability to secrete regenerative factors and modulate immune functions. However, issues remain regarding their further application for cell therapy. Here, to demonstrate the superiority of the improvement of MSCs, we divided umbilical cord blood-derived MSCs (UCB-MSCs) from 15 donors into two groups based on efficacy and revealed donor-dependent variations in the anti-inflammatory effect of MSCs on macrophages as well as their immunoregulatory effect on T cells. Through surface marker analyses (242 antibodies), we found that HLA-A2 was positively related to the anti-inflammatory and immunoregulatory function of MSCs. Additionally, HLA-A2 mRNA silencing in MSCs attenuated their therapeutic effects in vitro; namely, the suppression of LPS-stimulated macrophages and phytohemagglutinin-stimulated T cells. Moreover, HLA-A2 silencing in MSCs significantly decreased their therapeutic effects in a rat model of hyperoxic lung damage. The present study provides novel insights into the quality control of donor-derived MSCs for the treatment of inflammatory conditions and diseases.

Transplantation of umbilical cord blood-derived mesenchymal stem cells as therapy for adriamycin induced-cardiomyopathy

ABSTRACT Umbilical cord blood-derived mesenchymal stem cells (UCBMSCs) have been reported to possess cardioprotective effects in diseases. However, its effects on cardiomyopathy remain unclear. This study aimed to the therapeutic effects of UCBMSC transplantation on adriamycin (ADR)-induced cardiomyopathy. UCBMSCs isolated from human UCB were identified by detecting surface markers (CD29, CD90, CD34, and CD45) using flow cytometry. The effect of UCBMSCs on left ventricular end-diastolic dimension (LVEDD), left ventricular systolic end-diastolic diameter (LVESD), left ventricular ejection fraction (LVEF), and left ventricular fraction shortening (LVFS) were determined by echocardiography. Histological changes were observed by HE and Masson staining. The serum levels of collagen-I (Col-I), brain natriuretic peptide (BNP), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), creatine kinase (CK), CK-MB, interleukin (IL)-6, IL-10, and tumor necrosis factor alpha (TNF-α) were measured by corresponding kits. The protein levels of IL-6, IL-10, and TNF-α were measured by Western blotting. The isolated UCBMSCs manifested the positive expression of CD29 and CD90, and the negative expression of CD34 and CD45. UCBMSC transplantation significantly reduced LVEDD and LVESD, and increased LVEF and LVFS in ADR-induced cardiomyopathy model rats. Cardiac injury and high collagen deposition in model rats were alleviated by UCBMSC treatment. Moreover, UCBMSCs decreased the serum levels of Col-I, BNP, AST, LDH, CK, CK-MB, IL-6, IL-10, and TNF-α in model rats. Overall, UCBMSCs exert the therapeutic effects on ADR-induced cardiomyopathy through recovering the myocadiac function and alleviating the inflammatory response.

Immunomodulatory Effect of Epidermal Growth Factor Secreted by Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells on Atopic Dermatitis.

Background and ObjectivesHuman mesenchymal stem cells (MSCs) are emerging as a treatment for atopic dermatitis (AD), a chronic inflammatory skin disorder that affects a large number of people across the world. Treatment of AD using human umbilical cord blood-derived MSCs (hUCB-MSCs) has recently been studied. However, the mechanism underlying their effect needs to be studied continuously. Thus, the objective of this study was to investigate the immunomodulatory effect of epidermal growth factor (EGF) secreted by hUCB-MSCs on AD.Methods and ResultsTo explore the mechanism involved in the therapeutic effect of MSCs for AD, a secretome array was performed using culture medium of hUCB-MSCs. Among the list of genes common for epithelium development and skin diseases, we focused on the function of EGF. To elucidate the effect of EGF secreted by hUCB-MSCs, EGF was downregulated in hUCB-MSCs using EGF-targeting small interfering RNA. These cells were then co-cultured with keratinocytes, Th2 cells, and mast cells. Depletion of EGF disrupted immunomodulatory effects of hUCB-MSCs on these AD-related inflammatory cells. In a Dermatophagoides farinae-induced AD mouse model, subcutaneous injection of hUCB-MSCs ameliorated gross scoring, histopathologic damage, and mast cell infiltration. It also significantly reduced levels of inflammatory cytokines including interleukin (IL)-4, tumor necrosis factor (TNF)-α, thymus and activation-regulated chemokine (TARC), and IL-22, as well as IgE levels. These therapeutic effects were significantly attenuated at all evaluation points in mice injected with EGF-depleted hUCB-MSCs.ConclusionsEGF secreted by hUCB-MSCs can improve AD by regulating inflammatory responses of keratinocytes, Th2 cells, and mast cells.

Combined Mesenchymal Stem Cellsand Cartilage Acellular Matrix InjectionTherapy for Osteoarthritis in Goats.

Human umbilical cord blood-derived MSCs (hUCB-MSCs) have been studied in osteoarthritis (OA) and cartilage regeneration. Our previous study demonstrated that hUCB-MSCs combined with cartilage acellular matrix injection (CAM Inj.) represent potential therapeutic agents for structural improvement and anti-inflammatory effects in a rabbit model of OA. Based on a previous study, this study has evaluated the safety and efficacy of hUCB-MSCs combined with CAM Inj. in an anterior cruciate ligament transection (ACLT) with medial meniscectomy (MMx) in a goat model. In this study, 27 goats were divided into 5 groups: normal (n = 3), OA (n = 6), OA + CAM Inj. (n = 6), OA + hUCB-MSCs (n = 6), and OA + hUCB-MSCs + CAM Inj. (n = 6). Lameness and radiographic parameters were assessed 6months after administration, and macroscopic and histological evaluations of the goat articular cartilage were performed 6months after intervention. The results showed significant improvement in lameness score only in the OA + hUCB-MSCs group at 5months after treatment (*p < 0.05), whereas the K&L score showed significant improvement only in the OA + hUCB-MSCs + CAM Inj. group 6months after intervention (*p < 0.05). In addition, the gross findings showed significance in OA + CAM Inj. and OA + hUCB-MSCs + CAM Inj. groups 6months after treatment (*p < 0.05 and **p < 0.01). In conclusion, treatment with a combination of hUCB-MSCs and CAM Inj. reduced OA symptoms and induced effective cartilage tissue repair in a goat model. We suggest the combination of hUCB-MSCs and CAM Inj. as an alternative therapy for OA.

HYPERGLYCEMIA REVERSAL IN DIABETIC INFARCTED RAT POSTINTRAVENOUS INFUSION OF HUMAN MESENCHYMAL STEM CELLS

INTRODUCTION Hyperglycemia reversal and preservation/restoration of β-cells function in diabetic infarction remains as an attractive and challengeable therapeutic target. Mesenchymal stem cells (MSCs) are multipotent cells with a strong immunoregulatory potential that have emerged as a possible cell-based therapy for a variety of immunological diseases. The objective of this study was to examine the dose-dependent efcacy of intravenous administration of human umbilical cord blood derived MSCs (UCB-MSCs) in chemically induced rats with diabetic infraction. METHODS Wister rats (weight: 200-250g, males) received intraperitoneal streptozotocin injection followed by isoproterenol to develop diabetes infarction condition. After model development animals received intravenous single or double dose of human 6 UCB-MSCs (5 X 10 cells per animal at each dose) and followed up to 30 days post-administration. Pancreatic tissue histology, blood glucose and insulin levels were measured, and proportion of animal survival was calculated using Kaplan-Meier curve analysis. RESULTS Double dose of MSCs infusion resulted in reorganization of islet cells and partial restoration of β-cells at day 30. Comparatively faster restoration of glucose and insulin normalization was observed for two MSCs doses compared to single dose. Highest proportion of animal survival was observed (&gt;85%) for double doses of MSCs infusion compared to single dose (&gt;70%) at day 30. CONCLUSION Two consecutive intravenous doses of human UCB-MSCs can improve structural and functional decits of pancreatic tissues and maintain blood glucose and insulin levels in diabetic infarcted rats up to 30 days. However, identication of long-term effects entails longer follow-up periods, and larger sample sizes with other investigations.

Immunomodulatory Effect of Human Umbilical Cord Blood-derived Mesenchymal Stem Cells on Activated T-lymphocyte.

Many studies have been performed about regenerative and immunomodulatory properties of mesenchymal stem cells (MSCs) and their application in different treatment approaches. The present study aimed to investigate the immunomodulatory effect of umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) on the gene expression profile of cytokines in stimulated T-lymphocytes. For this purpose, MSCs were isolated from umbilical cord blood samples and cultured in Dulbecco's Modified Eagle Medium supplemented with 10% fetal bovine serum. The nature of MSCs was identified by flow cytometry analysis and differentiation to the adipocyte and osteocyte lineage. Moreover, to investigate the immunomodulatory effects of MSCs on T cells, a co-culture system was designed and expression levels of interleukin (IL)-2, IL-4, IL-6, IL-10,IL-13, interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), and transforming growth factor-beta (TGF-β) genes were measured; using the real-time polymerase chain reaction (RT-PCR) technique. Our results demonstrated the ability of MSCs to differentiate into adipocyte and osteocyte lineages. Further investigation also displayed that although UCB-MSCs could significantly reduce the expression of pro-inflammatory cytokines like IL-2, IL-6, IFN-γ, and TNF-α in activated T-lymphocytes, they noticeably potentiated the expression levels of IL-4, IL-10, IL-13, and TGF-β in the co-culture setting. In conclusion, UCB-MSCs have immunomodulatory effects on activated T-lymphocytes in favor of anti-inflammatory responses.

158&#x2003;Umbilical cord blood-derived mesenchymal stem cells (UCB-MSC) used for the prevention of metritis in cattle.

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AACE2021-A-1055: Efficacy of Umbilical Cord Blood-Derived Stem Cells Transplantation Versus Their Conditioned Media in Hypothyroid Rats on Thyroid Function and Structure

Primary hypothyroidism is a chronic condition with significant morbidity. Providing physiological thyroid hormone replacement through pharmacological treatment remains controversial. Aim of work: To assess the efficacy of human umbilical cord blood-derived mesenchymal stem cells (UM-MSCs) transplantation versus their conditioned media (CM) on thyroid function tests (TFTs) and thyroid structure in hypothyroid rats, also to compare intravenous versus its intranasal route.

LINC02381, a sponge of miR-21, weakens osteogenic differentiation of hUC-MSCs through KLF12-mediated Wnt4 transcriptional repression

Human umbilical cord blood-derived MSCs (hUC-MSCs) have the potential to differentiate into osteoblasts. This study investigated the function and potential mechanisms of a novel lncRNA LINC02381 in hUC-MSC osteogenic differentiation. hUC-MSCs were maintained in osteogenic differentiation medium. RT-qPCR assay was performed to assess LINC02381 expression. Alizarin Red S (ARS) and alkaline phosphatase (ALP) staining were performed to evaluate osteogenic differentiation. The interaction between miR-21 and LINC0238/KLF12 was determined by luciferase reporter and RNA immunoprecipitation (RIP) assays. Chromatin immunoprecipitation (ChIP) assay was used to confirm the transcriptional regulation of KLF12 on Wnt4 promoter. The nuclear translocation of β-catenin was evaluated using immunofluorescence. hUC-MSCs seeded on Bio-Oss Collagen scaffolds were transplanted into nude mice to assess in vivo osteogenesis. Bone formation was observed by H&E and Masson's trichrome staining. OSX and OPN levels were assessed by immunohistochemistry. LINC02381 was up-regulated in the clinical samples of osteoporotic patients. However, LINC02381 expression was reduced during osteogenic differentiation of hUC-MSCs. Enforced expression of LINC02381 suppressed the osteogenic differentiation of hUC-MSCs. Mechanistically, LINC02381 sponged miR-21 to enhance KLF12 expression, which led to the inactivation of Wnt/β-catenin signaling pathway. Furthermore, miR-21 mimics or KLF12 silencing counteracted LINC02381-induced inhibition of osteogenic differentiation, whereas IWP-4 (an inhibitor of Wnt pathway) abolished this effect. In summary, LINC02381 repressed osteogenic differentiation of hUS-MSCs through sponging miR-21 to enhance KLF12-mediated inactivation of Wnt/β-catenin pathway, indicating that LINC02381 might be a therapeutic target for osteoporosis.