Human Umbilical Cord Blood Mesenchymal Stem Cells Research Articles

The combined effect of the human umbilical cord blood with chitosan scaffold on the full-thickness wound healing process in rats

This studyaimed to investigate the role of a low dose of human umbilical cord blood mesenchymal stem cells (HUCB-MSCs) with chitosan scaffold (ChSc) on the progress of the cutaneous wound healing process in male Albino rats. A full-thickness cutaneous wound with a circular shape with a diameter of about 2 cm was induced on the dorsum of thirty-six Albino male rats, divided into four groups: the first group, the wounds were left without treatment as a control group. In the second group, the wounds were covered by freeze-dried ChSc. While in the third group, the wounds were treated through injection of the HUCB-MSCs intradermally, and in the fourth group, the ChSc seeded with HUCB-MSCs were used together to treat the wounds. The progress of wound healing was monitored by histological, immunohistochemical, and biochemical assays for all experimental groups at 3, 8, and 21 postoperative days. Both ChSc and the low dose of HUCB-MSCs alone performed moderate healing progress.The (HUCB-MSCs) ChSc group exhibited an increased healing rate more than the other groups and reported an appropriate collagen deposition without scarring signs, effective mast cell regulation, and well vascularization. In conclusion, the fourth group (the HUCB-MSCs with ChSc) improved the healing process, revealing the highest healing rate and performance without complications.

3D-printed collagen/silk fibroin/secretome derived from bFGF-pretreated HUCMSCs scaffolds enhanced therapeutic ability in canines traumatic brain injury model.

The regeneration of brain tissue poses a great challenge because of the limited self-regenerative capabilities of neurons after traumatic brain injury (TBI). For this purpose, 3D-printed collagen/silk fibroin/secretome derived from human umbilical cord blood mesenchymal stem cells (HUCMSCs) pretreated with bFGF scaffolds (3D-CS-bFGF-ST) at a low temperature were prepared in this study. From an in vitro perspective, 3D-CS-bFGF-ST showed good biodegradation, appropriate mechanical properties, and good biocompatibility. In regard to vivo, during the tissue remodelling processes of TBI, the regeneration of brain tissues was obviously faster in the 3D-CS-bFGF-ST group than in the other two groups (3D-printed collagen/silk fibroin/secretome derived from human umbilical cord blood mesenchymal stem cells (3D-CS-ST) group and TBI group) by motor assay, histological analysis, and immunofluorescence assay. Satisfactory regeneration was achieved in the two 3D-printed scaffold-based groups at 6 months postsurgery, while the 3D-CS-bFGF-ST group showed a better outcome than the 3D-CS-ST group.

VEGF165 gene-modified human umbilical cord blood mesenchymal stem cells protect against acute liver failure in rats.

Human umbilical cord blood mesenchymal stem cells (HUCB-MSCs) can exert a protective effect in rat models of acute liver failure (ALF). Vascular endothelial growth factor 165 (VEGF165 ) is the predominant VEGF isoform and possesses a strong pro-angiogenic function. In the present study, HUCB-MSC served as the gene delivery vehicle for the VEGF165 gene, and we explored the therapeutic effects of this system on ALF. HUCB-MSCs were infected with an adenovirus expressing green fluorescent protein (GFP)-VEFG fusion protein (Ad-VEGF165 ) to overexpress VEGF165 or an adenovirus expressing GFP (Ad-GFP) as control. The control and modified HUCB-MSCs were then transplanted into ALF model rats. Liver function and liver pathological changes were assessed by biochemical tests and liver histology. Immunohistochemistry was carried out to determine the expression of, CD34, Ki67 and VEGF. VEGF165 overexpression enhanced the multipotency of HUCB-MSCs and promoted the homing and colonization of HUCB-MSC in the liver tissues of ALF rats. Furthermore, although HUCB-MSC transplantation ameliorated liver damage and promoted liver regeneration to some extent in ALF rats, Ad-VEGF165 -HUCB-MSC transplantation showed stronger therapeutic effects on ALF. In summary, transplantation of VEGF165 -modified HUCB-MSCs exert stronger therapeutic effects on ALF than HUCB-MSCs. The present study provides a novel therapeutic approach for ALF.

Rapid Induction of Mesenchymal Stem Cells to Neural Cells

Background:-Human umbilical cord blood- mesenchymal stem cells(HUCB-MSCs) hold great promise invitro neural differentiation and therapy for neurodegenerative disorders. It has demonstrated that the numberand differentiating potential of bone marrow mesenchymal stem cells (MSCs) decrease with age. Therefore,the search for alternative sources of MSCs is of significant value.Objective:-To determine the possibility of obtaining clonally expanded UCB-MSCs and evaluate their rapidinduction and differentiation into neural cells in vitro.Methods:-MSCs were isolated from HUCB by combining gradient density centrifugation with plasticadherence. In order to identify factors that able to lead to neural differentiation, the cultured cells weretreated with retinoic acid (RA) and B-mercaptoethanol (BME). Differentiating characterization of UCBMSCswere detected by immunocytochemistry analysis.Results:-UCB-MSCs appeared like the fibroblast cell and these cells were extensively expanded in culture.The results showed that UCB-MSCs were positive for CD71and CD90 but were negative for CD34. Theimmunocytochemistry staining indicated that the differentiated cells give positive response for nestinemarker. The result confirmed for neural progenitor differentiation of MSCs.Conclusion: The results of this study confirmed that UCB provides a great source of stem cells for using intreatment of neurodegenerative disorders of the central nervous system.

Tropism of mesenchymal stem cell toward CD133+ stem cell of glioblastoma in vitro and promote tumor proliferation in vivo

BackgroundPrevious studies have demonstrated remarkable tropism of mesenchymal stem cells (MSCs) toward malignant gliomas, making these cells a potential vehicle for delivery of therapeutic agents to disseminated glioblastoma (GBM) cells. However, the potential contribution of MSCs to tumor progression is a matter of concern. It has been suggested that CD133+ GBM stem cells secrete a variety of chemokines, including monocytes chemoattractant protein-1 (MCP-1/CCL2) and stromal cell-derived factor-1(SDF-1/CXCL12), which could act in this tropism. However, the role in the modulation of this tropism of the subpopulation of CD133+ cells, which initiate GBM and the mechanisms underlying the tropism of MSCs to CD133+ GBM cells and their effects on tumor development, remains poorly defined.Methods/resultsWe found that isolated and cultured MSCs (human umbilical cord blood MSCs) express CCR2 and CXCR4, the respective receptors for MCP-1/CCL2 and SDF-1/CXCL12, and demonstrated, in vitro, that MCP-1/CCL2 and SDF-1/CXC12, secreted by CD133+ GBM cells from primary cell cultures, induce the migration of MSCs. In addition, we confirmed that after in vivo GBM tumor establishment, by stereotaxic implantation of the CD133+ GBM cells labeled with Qdots (705 nm), MSCs labeled with multimodal iron oxide nanoparticles (MION) conjugated to rhodamine-B (Rh-B) (MION-Rh), infused by caudal vein, were able to cross the blood-brain barrier of the animal and migrate to the tumor region. Evaluation GBM tumors histology showed that groups that received MSC demonstrated tumor development, glial invasiveness, and detection of a high number of cycling cells.ConclusionsTherefore, in this study, we validated the chemotactic effect of MCP-1/CCL2 and SDF-1/CXCL12 in mediating the migration of MSCs toward CD133+ GBM cells. However, we observed that, after infiltrating the tumor, MSCs promote tumor growth in vivo probably by release of exosomes. Thus, the use of these cells as a therapeutic carrier strategy to target GBM cells must be approached with caution.

Activated Schwann cells combined with umbilical cord blood mesenchymal stem cell transplantation for repair of spinal cord injury

Objective To observe the variation of neuronal markers after co-transplantation of activated Schwann cells (ASCs) and human umbilical cord blood mesenchymal stem cells (HUCBMSCs)into rat spinal cord injury (SCI). Also, we discuss whether these two types of cells grafted into the injured site of spinal cord have the ability of promoting axonal regeneration and functional recovery after SCI. Methods SCI model in Wistar rat (Female, 8 weeks’ old) was created at T10 segment with NYU Impactor-Ⅱmachine of 10 g×50 mm weight drop.80 SCI rats were classified into 4 groups randomly: blank control group, ASCs transplantation group, HUCBMSCs transplantation group and co-transplantation group of ASCs and HUCBMSCs. 1, 2, 3, 4 weeks after operation, 3 animals of each group were sacrificed, and injured spinal cord tissue were used for Western blotting and Real-time polymerase chain reaction analysis [nuclear factor (NF)-200, glial fibrillary acidic protein (GFAP), myelin basic protein (MBP)]. Basso, Beattie, and Bresnahan (BBB) score was carried out at the 3rd day and at weekly after intervals injury.12 weeks later, 8 rats in each group were selected to carry out 10% BDA anterograde tracing mark.2 weeks later, the animals were sacrificed, the spinal cord tissue with injured lesion was taken out and fast cryostat sections (5 μm) was made followed by BDA developing, hematoxylin and eosin (HE) and immunochemistry (NF-200, MBP, GFAP) staining. Results Western blotting and Real-time PCR analysis revealed that NF-200 and MBP expressed higher with time while the co-graft group is the highest (4 weeks after injury, NF-200: 4.581±0.588, MBP: 3.922±0.259). From the 4th week after cell-transplantation, BBB score in co-graft group was significantly higher than that in other 3 groups (P=0.000). The rats of co-graft group recoverd to the largest extent. BDA anterograde injured site in co-graft group than that in other 3 groups, while little in control group (P=0.000). HE staining showed that the injured cavity was the smallest one in cell co-graft group when compared with other 3 groups. Conclusion The results indicate that the co-transplantation fully takes advantage of ASCs to promote HUCBMSCs differentiate into neurons. This is more effective than single cell transplantation in promoting axonal regeneration and functional recovery after rat’s spinal cord injury. Key words: Umbilical cord blood mesenchymal stem cells; Activated Schwann cells; Spinal cord injury; Neural repair; Differentiation; Co-transplantation

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.

The regenerative effect of human umbilical cord blood mesenchymal stem cells in a rabbit model of osteoarthritis

Osteoarthritis (OA) is a degenerative disorder characterized by changes in subchondral and periarticular bone. The limited number of therapeutic choices for articular injury and disease has increased the need for stem cells as a biological replacement for damaged cartilage. Umbilical cord (UC) blood cells are easily available and less immunogenic than other sources of stem cells, and there are no ethical concerns surrounding their use. These cells are isolated from young donors. Human umbilical cord blood mesenchymal stem cells (HUCB-MSCs) appear to be an ideal candidate for cartilage regeneration. The aim of the study was to investigate the regenerative effect of HUCB stem cells on degenerated articular cartilage in New Zealand white rabbits experimentally induced with OA. This study was performed on 42 New Zealand white rabbits. They were surgically induced with OA in the left knees by cutting the anterior cruciate ligament. After confirmation of the development of OA histopathologically, we delivered a single dose of HUCB-MSCs directly intra-articularly in the cell-treated groups. Controls were injected with only suspension media. Histopathological tests were performed 8 and 24 weeks after injection. Histopathologically, 8 weeks after the injection, cell-treated rabbits showed better cartilage quality and lower degree of degeneration, whereas 24 weeks after the injection all parameters in the cell-treated groups were significantly better. HUCB-MSCs are a promising stem cell source for cartilage tissue formation and can decrease the development of OA in rabbits.

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.

SAT0546 The Regenerative Effect of Human Umbilical Cord Blood Mesenchymal Stem Cells in A Rabbit Model of Osteoarthritis

SAT0546 The Regenerative Effect of Human Umbilical Cord Blood Mesenchymal Stem Cells in A Rabbit Model of Osteoarthritis

Impact of activated schwann cells-derived neuronotrophins on differentiation of human umbilical cord blood mesenchymal stem cells into neurogenic cellsin vitro

Objective To observe the capability of human umbilical cord blood mesenchymal stem cells (HUCBMSCs) to differentiate into neurogenic cells in vitro under the impact of activated Schwann cells (ASCs)-derived neuronotrophins.Methods ASCs were cultured by using the dual-enzyme digestion method combined with mechanical separation.HUCBMSCs were cultured through isolating HCMNCs by Ficoll' s way and then plastic-adherent culture.Two methods were used for HUCBMSCs culture:all-trans retinoic acid + bFGF + EGF ( control group),1/2 DMEM + ASCs culture ( medium after ASCs cultured for 2 days,experimental group).Immunohistochemistry,Western blotting,and semi-quantitative and realtime polymerase chain reaction (PCR,NF200,GFAP,MBP) quantitative analysis were used to evaluate neurogenic differentiation of HUCBMSCs.Results The results of immunohistochemistry staining,such as NF200,GFAP and MBP,confirmed that HUCBMSCs could differentiate into neurogenic cells under the effect of ASCs and RA + bFGF + EGF.The statistical analysis of Western blotting and RT-PCR ( NF200,GFAP,MBP ) indicated that there existed significant difference between two groups.Conclusion HUCBMSCs can proliferate and differentiate into neurogenic cells under the effect of numerous neurotrophic factors from ASCs in vitro. Key words: Umbilical cord blood mesenchymal stem cells; Schwann cells; Neural repair; Differentiation

In vitro labeling and MRI of mesenchymal stem cells from human umbilical cord blood

The aim of this study was to label human umbilical cord blood mesenchymal stem cells (MSCs) with poly-l-lysine (PLL)-conjugated superparamagnetic iron oxide particles and to obtain magnetic resonance (MR) images of the labeled MSCs' suspension at 1.5 T. PLL was conjugated with iron oxide to form superparamagnetic particles called Fe(2)O(3)-PLL. Human umbilical cord blood MSCs were isolated, purified, expanded and incubated with Fe(2)O(3)-PLL. Prussian blue stain was performed to show intracellular iron; spectrometry was used to quantify iron uptake within cells. Tetrazolium salt (MTT) assay was applied to evaluate toxicity and proliferation of MSCs labeled with various concentrations of Fe(2)O(3)-PLL. The cell apoptosis rate was determined by annexin V/propichium iodide (PI) double staining method. Vials containing cells underwent MR imaging (MRI) with T(1), T(2) and T(2)* weighted MRI. Iron-containing intracytoplasmatic vesicles could be observed clearly with Prussian blue staining in all samples except the unlabeled control. The iron content per cell determined by spectrometry was 64.51+/-10.32 pg. Among MSCs with and without labeling of various concentrations of Fe(2)O(3)-PLL, MTT values of light absorption had no statistically significant difference (Kruskal-Wallis test, chi(2)=10.35, P=.17). A concentration at 20 mug/ml of iron appeared most suitable for incubating cells. Of labeled and unlabeled MSCs, the early [annexin V-fluorescein isothiocyanate (FITC)-positive/PI-negative] and late (annexin V-FITC-positive/PI-positive) apoptotic cells were 10.34+/-0.43%/11.36+/-1.30% and 4.01+/-1.76%/2.98+/-1.37%, respectively, and there were no significant differences between them (P>.05). T(2) weighted image (WI) and T(2)*WI demonstrated significant decrease of signal intensity (SI) in vials containing 1 x 10(6) (1 day), 1x10(6) (8 days) and 5 x 10(5) labeled cells, in comparison with unlabeled cells (P<.05). The percentage change of SI (DeltaSI) was significantly higher in 10(6) labeled cells after 1-day culture than that in the same number of labeled cells after 8-day culture and that in 5 x 10(5) labeled cells, particularly on T(2)*WI (P<.05). Among pulse sequences, T(2)*WI demonstrated the highest DeltaSI (P<.05). The human umbilical cord blood MSCs can be labeled with Fe(2)O(3)-PLL without significant change in viability and apoptosis. The suspension of labeled MSCs can be imaged with standard 1.5-T MR equipment.

Induction of umbilical cord blood mesenchymal stem cells into neuron-like cells in vitro.

Mesenchymal stem cells (MSCs) in human umbilical cord blood are multipotent stem cells that differ from hematopoietic stem cells. They can differentiate in vitro into mesenchymal cells such as osteoblasts and adipocytes. However, differentiation into nonmesenchymal cells has not been demonstrated. Here, we report the isolation, purification, expansion, and differentiation of human umbilical cord blood MSCs into neurocytes in vitro. Cord blood samples were allowed to drain from the end of the cord into glass bottles with 20 U/mL preservative-free heparin. MSCs were isolated from human umbilical cord blood, purified, and expanded in Mesencult medium. Surface antigens of MSCs were analyzed by fluorescence-activated cell sorting (FACS). MSC passages 2,5, and 8 were induced to differentiate into neuron-like cells. Neurofilament (NF) and neuron-specific enolase (NSE) were detected by immunohistochemistry staining. Special Nissl bodies were observed by histochemical analysis. The results showed that 6.6 x 10(5) primary MSCs were expanded for 10 passages to obtain 9.9 x 10(8) MSCs, an increase of approximately 1.5 x 10(3)-fold. FACS results showed that the MSCs did not express antigens CD34, CD11a, and CD11b and expressed CD29 and CD71, an expression pattern identical to that of human bone marrow-derived MSCs. Induction results indicated that approximately 70% of the cells exhibited a typical neuron-like phenotype. Immunohistochemistry staining suggested that induced MSCs of different passages expressed NF and NSE. Special Nissl bodies were obvious in the neuron-like cells. These results suggest that MSCs in human umbilical cord blood are capable of differentiating into neuron-like cells in vitro.