骨髓间充质干细胞促进“创面”愈合的实验研究

目的：探讨骨髓间充质干细胞（MSCs）促进创面愈合的作用和用于创面修复的可行性。方法：密度梯度离心法分离培养正常人骨髓间充质干细胞（hMSCs），流式细胞仪检测培养第二代细胞CD14、CD29、CD34、CD44、CD45和CD106抗原表达以鉴定MSCs。实验分3组：直接共培养组（n=8）为hMSCs（1×10^5/ml）与人表皮角质形成细胞（HEKa）直接接触共培养；间接共培养组（n=7）为在transwell透明聚碳酸酯膜上层加入hMSCs细胞悬液[（2-3）×10^5个细胞）]，下层接种HEKa，间接共培养；单纯HEKa培养组（n=7）单纯培养HEKa。在倒置显微镜下刮擦生长融合成片的HEKa细胞以制备宽度为100μm的“表皮创面”模型。模型制备后24、48、72h，倒置相差显微镜下计数每高倍视野越过创缘迁移到创面的HEKa数，并用SigmaScan Pro5软件计算创面愈合率，检测HEKa细胞吸收脱氧胸腺嘧啶核苷的放射活性，判定HEKa细胞的分裂增殖活性。结果：流式细胞仪检测第二代hMSCs CD29、CD44和CD106抗原阳性（分别为94.81%、95.38%、97.40%），CD14、CD34、CD45抗原阴性（分别为1.23%、3.05%、2.64%）；直接培养组、间接培养组和单纯HEKa培养组越过创缘进入创面的细胞伤后24h分别有（10.00±0.25）、（5.50±0.20）和（5.20±0.35）个/高倍视野；48h分别有（55.30±0.22）、（27.00±0.34）和（26.50±0.25）个/高倍视野；72h分别有（110.50±0.45）、（42.50±0.50）、（40.20±0.38）个/高倍视野。3组伤后72h创面愈合率分别为（60.00±0.04）%，（35.00±0.01）%、（33.00±0.05）%。脱氧胸腺嘧啶苷掺入培养基法表明hMSCs对体外共培养HEKa细胞的增殖作用分别为（1650±270）cpm/10^5 cell、（1240±210）cpm/10^5 cell、（1180±220）cpm/10^5 cell。上述各指标直接共培养组与单纯HEKa培养组比较，差异有显著性（P〈0.01），而间接培养组与单纯HEKa培养组间差异无显著性（P〉0.05）。     

Mesenchymal stem cells combined with an artificial dermal substitute improve repair in full-thickness skin wounds

Autografts represent the gold standard for the treatment of full thickness burns. Factors such as lack of suitable donor sites and poor skin quality, however, have led to the development of artificial dermal substitutes. The investigation of mechanisms leading to enhanced functionality of these skin substitutes has been attracting great attention. This study aimed to investigate the effect of autologous stem cells on the integration and vascularization of a dermal substitute in full-thickness skin wounds, in a murine model. Two cell populations were compared, whole bone marrow cells and cultivated mesenchymal stem cells, isolated from mice transgenic for the enhanced green fluorescent protein, which allowed tracking of the transplanted cells. The number of cells colonizing the dermal substitute, as well as vascular density, were higher in mice receiving total bone marrow and particularly mesenchymal stem cells, than in control animals. The effect was more pronounced in animals treated with mesenchymal stem cells, which located primarily in the wound bed, suggesting a paracrine therapeutic mechanism. These results indicate that combining mesenchymal stem cells with artificial dermal substitutes may represent an important potential modality for treating full thickness burns, even in allogeneic combinations due to the immunoregulatory property of these cells.     

Evaluation of autologous bone marrow‐derived nucleated cells for healing of full‐thickness skin wounds in rabbits

The aim of the study was to evaluate the potential of autologous bone marrow‐derived nucleated cells to enhance the rate of healing of full‐thickness excisional skin wounds in rabbits. The study was conducted on 20 New Zealand white rabbits of either sex. Two, 2 × 2 cm full‐thickness skin (thoracolumabar region) excisional wounds were created; one on each side of the dorsal midline in each animal. The wounds were randomly assigned to either injection of autologous bone marrow‐derived nucleated cells into the wound margins (BI), or topical application of sterile saline solution (normal saline, NS), which served as control. The wound healing was assessed by evaluation of granulation tissue formation, wound contraction, epithelisation and histopathological and histochemical changes up to 28 days after creation of the wound. Granulation tissue appeared significantly faster in BI‐treated wounds (3.22 ± 0.22 days) than in NS‐treated wounds (4.56 ± 0.47 days). Better epithelisation was seen histologically in BI wounds than in NS‐treated wounds. Wound contraction was significantly more in BI wounds when compared with NS wounds on 21 post‐surgery. Histopathological examination of the healing tissue showed early disappearance of inflammatory reaction, significantly more neovascularisation, and more fibroplasias and early lay down and histological maturation of collagen in BI wounds than in control wounds. It was concluded that injection of autologous bone marrow‐derived nucleated cells in the wound margins induced faster and better quality healing of excisional skin wounds in rabbits when compared with normal saline. The injection of autologous bone marrow‐derived nucleated cells can be used to promote healing of large full‐thickness skin wounds in rabbits.     

Development and characterization of novel clinical grade neonatal porcine bone marrow‐derived mesenchymal stem cells

Due to recent advances in research on mesenchymal stem cells (MSCs), MSCs are expected to be used in various clinical applications. However, securing adequate cadaveric donors and safety of living donors are major issues. To solve such issues, we have examined to develop clinical grade neonatal porcine bone marrow‐derived MSCs (npBM‐MSCs). Clinical grade neonatal porcine bone marrow cells were collected, frozen, and sent to our laboratory by air. The npBM‐MSCs were isolated from thawed bone marrow cells, then frozen. The thawed npBM‐MSCs were examined for CD markers and differentiated into chondrocytes, osteocytes, and adipocytes. They were compared with human bone marrow‐derived MSCs (hBM‐MSCs) for growth rate and size. To assess the robustness of proliferation, we compared culture medium with or without gelatin. The npBM‐MSCs expressed positive MSC markers CD29, CD44, and CD90 and were differentiated into chondrocytes, osteocytes, and adipocytes. The doubling time of npBM‐MSCs was significantly shorter than that of hBM‐MSCs (17.3 ± 0.8 vs 62.0 ± 19.6 hours, P < 0.01). The size of npBM‐MSCs was also significantly smaller than that of hBM‐MSCs (13.1 ± 0.3 vs 17.5 ± 0.4 μm, P < 0.001). The npBM‐MSCs showed similar proliferation characters irrespective of with or without gelatin coating. The npBM‐MSCs secreted VEGF‐A, VEGF‐C, and TGF‐β1. We have established npBM‐MSCs which show super‐rapid growth, small size, and robust proliferation profile. The np‐MSCs might be able to solve the donor issues for MSC therapy.     

Autologous bone‐marrow‐derived mesenchymal stem cell transplantation into injured rat urethral sphincter

Objectives: To evaluate the functional and histological recovery by autologous bone‐marrow‐derived mesenchymal stem cell (BMSC) transplantation into injured rat urethral sphincters. Methods: BMSC were harvested from female Sprague–Dawley retired breeder rats for later transplantation. The cells were cultured, and transfected with the green fluorescence protein gene. The urethral sphincters were injured by combined urethrolysis and cardiotoxin injection. One week after injury, the cultured BMSC were injected autologously into the periurethral tissues. Controls included sham‐operated rats and injured rats injected with cell‐free medium (CFM). Abdominal leak point pressures (LPP) were measured before and after surgery during the following 13 weeks. The urethras were then retrieved for histological evaluation. The presence of green‐fluorescence‐protein‐labeled cells and the regeneration of skeletal muscles, smooth muscles, and peripheral nerves were evaluated by immunohistochemical staining. Results: LPP was significantly reduced in the injured rats. It increased gradually after transplantation, but there was no significant difference between the BMSC and CFM groups. In the BMSC group, transplanted cells survived and differentiated into striated muscle cells and peripheral nerve cells. The proportions of skeletal muscle cells and peripheral nerves in the urethra were significantly greater in the BMSC group compared to the CFM group. Conclusions: Despite a clear trend towards recovery of LPP in BMSC‐transplanted urethras, no significant effect was detected. Further study is required for clinical applications for the treatment of stress urinary incontinence.     

Efficacy of mesenchymal stem cell-delivery using perpendicular multi-needle injections to the skin: Evaluation of post-ejection cellular health and dermal delivery

AimMesenchymal stem cell (MSC)-therapy is increasingly being evaluated in clinical trials. Dermal delivery is not only time consuming but also unreliable, potentially hampering the therapeutic result. Therefore, qualification of cell delivery protocols is essential. This study evaluated a clinically relevant automated multi-needle injection method for cutaneous MSC-therapy, allowing the skin to be readily and timely treated, by assessing both the cellular health post-ejection and dermal delivery.MethodsFollowing dispensation through the injector (31 G needles: 9- or 5-pin) the cellular health and potency (perceived- and long-term (12 h) viability, recovery, metabolism, adherence, proliferation and IDO1-expression) of adipose-derived stem cells (10–20–50 ×10⁶ cells/ml) were assessed in vitro in addition to dermal delivery of solution in human skin.ResultsNo significant detrimental effect on the perceived cell viability, recovery, metabolism, adherence or IDO1-expression of either cell concentration was observed. However, the overall long-term viability and proliferation decreased significantly regardless of cell concentration, nonetheless marginally. An injection depth above 1.0 mm resulted in all needles piercing the skin with dermal delivery from up to 89% needles and minimal reflux to the skin surface, and the results were confirmed by ultrasound and histology.ConclusionThe automated injector is capable of delivering dermal cell-doses with an acceptable cell quality.     

Mobilised bone marrow‐derived cells accelerate wound healing

Massive skin defects caused by severe burn and trauma are a clinical challenge to surgeons. Timely and effective wound closure is often hindered by the lack of skin donor site. Bone marrow‐derived cells (BMDCs) have been shown to ‘differentiate’ into multiple tissue cells. In this study we focused on the direct manipulation of endogenous BMDCs, avoiding the immunocompatibility issues and complicated cell isolation, purification, identification and amplification procedures in vitro on wound repair. We found that mobilisation of the BMDCs into the circulation significantly increased the amount of BMDCs at the injury site which in turn accelerated healing of large open wound. We used a chimeric green fluorescent protein (GFP) mouse model to track BMDCs and to investigate their role in full‐thickness skin excisional wounds. We have shown that bone marrow mobilisation by granulocyte colony stimulating factor (G‐CSF) exerted multiple beneficial effects on skin repair, both by increasing the engraftment of BMDCs into the skin to differentiate into multiple skin cell types and by upregulating essential cytokine mRNAs critical to wound repair. The potential trophic effects of G‐CSF on bone marrow stem cells to accelerate wound healing could have a significant clinical impact.     

Mesenchymal stem cell therapy of acute thermal burns: A systematic review of the effect on inflammation and wound healing

AimMesenchymal stem cell (MSC) therapies are emerging as a promising strategy to promote tissue repair, and may extend their utility to burn care. This comprehensive review of the extant literature, evaluated all in vivo studies, to elucidate the potential protective and therapeutic effect of MSCs in acute thermal skin burns.MethodsPubMed was systematically searched, according to PRISMA guidelines, and all relevant preclinical and clinical studies were included according to pre-specified eligibility criteria.ResultsForty-two studies were included in a qualitative synthesis, of which three were human and 39 were animal studies. The preclinical studies showed that MSCs can significantly reduce inflammation, burn wound progression and accelerate healing rate of acute burns. The underlying mechanisms are complex and not fully understood but paracrine modulators, such as immunomodulatory, antioxidative and trophic factors, seem to play important roles. Allogeneic MSC therapy has proved feasible in humans, and could allow for prompt treatment of acute burns in a clinical setting.ConclusionMSC therapy show positive results, regarding improved burn wound healing and immunologic response. However, most findings are based on small animal studies. Randomized clinical trials are warranted to investigate the regenerative effects in human burns before translating the findings into clinical practice.     

人增生性瘢痕成纤维细胞的间充质干细胞表型及多向分化潜能

目的 探讨人增生性瘢痕来源的成纤维细胞的细胞表型及分化潜能,以进一步研究其在增生性瘢痕形成中的作用.方法 从手术切除的增生性瘢痕中提取并分离培养成纤维细胞,以倒置显微镜观察其细胞形态及生长特点.待细胞培养传代至第3代,采用Vi-CELL细胞计数仪检测细胞生长情况,流式细胞仪检测成纤维细胞间充质干细胞表型标志CD73、CD44、CD105、CD90、CD34、CD45、CD14的表达情况;通过免疫细胞化学检测该细胞CK19、Oct 4、Vementin的表达情况及免疫荧光检测α-平滑肌肌动蛋白;诱导分化检测细胞向成骨、成软骨和成脂细胞方向分化能力.结果 细胞原代培养初期贴壁散在分布,生长曲线显示细胞1 ～2d生长较慢,从3～5d左右细胞生长较快,6～7d细胞进入平台期;第2代第5天细胞多为长梭形,呈放射状、漩涡状排列.细胞流式结果显示细胞表型CD90、CD44、CD105、CD73呈高表达,CD45、CD34、CD14不表达;免疫细胞化学检测间充质细胞标记物Vementin、多能干细胞标志物Oct 4均呈阳性表达,免疫荧光检测α-平滑肌肌动蛋白呈阳性表达,上皮细胞标志物CK19呈阴性表达,多向诱导分化实验该细胞可向成骨、成软骨和成脂细胞分化,具有多向分化潜能.结论 人增生性瘢痕来源成纤细胞具有间充质干细胞样生物学特性,该生物学特性可能在增生性瘢痕的形成以及创面修复中发挥至关重要的作用.     

Comparison of the osteogenic capacity of minipig and human bone marrow‐derived mesenchymal stem cells

Minipigs are a recommended large animal model for preclinical testing of human orthopedic implants. Mesenchymal stem cells (MSCs) are the key repair cells in bone healing and implant osseointegration, but the osteogenic capacity of minipig MSCs is incompletely known. The aim of this study was to isolate and characterize minipig bone marrow (BM) and peripheral blood (PB) MSCs in comparison to human BM‐MSCs. BM sample was aspirated from posterior iliac crest of five male Göttingen minipigs (age 15 ± 1 months). PB sample was drawn for isolation of circulating MSCs. MSCs were selected by plastic‐adherence as originally described by Friedenstein. Cell morphology, colony formation, proliferation, surface marker expression, and differentiation were examined. Human BM‐MSCs were isolated and cultured from adult fracture patients (n = 13, age 19–60 years) using identical techniques. MSCs were found in all minipig BM samples, but no circulating MSCs could be detected. Minipig BM‐MSCs had similar morphology, proliferation, and colony formation capacities as human BM‐MSCs. Unexpectedly, minipig BM‐MSCs had a significantly lower ability than human BM‐MSCs to form differentiated and functional osteoblasts. This observation emphasizes the need for species‐specific optimization of MSC culture protocol before direct systematic comparison of MSCs between human and various preclinical large animal models can be made. © 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 30:1019–1025, 2012     

Insights into bone marrow‐derived mesenchymal stem cells safety for cutaneous repair and regeneration

Wound healing involves the orchestration of a complex process of interactions between numerous types of cell, components of extracellular matrix and signalling molecules following injury, which is usually a highly successful biological course to reconstruct the integrity of the skin. Nevertheless, when skin is severely damaged, the injured skin is limited in its ability to repair itself and possibly results in the hypertrophic scars or so‐called keloids, and non healing wound or ulcer. Bone marrow‐derived mesenchymal stem cells (BM‐MSCs) are being clinically explored as a promising therapy in the field of tissue repair and regeneration. However, potential risks associated with these cell‐based therapies remain uncertain. The aim of this review is to summarise the safety issues accompanying the administration of BM‐MSCs for acute or chronic skin repair and regeneration. More importantly, this review highlights the requirement for fundamental research to improve future clinical application of these strategies, as well as for regulatory authorities to establish clinical criteria to identify the qualitative requirements for the manufacture process of cells products, which will ensure the manufacture process of the best benefit‐to‐risk ratio of cell‐based therapy for the patients.     

Isolation and characterization of turkey bone marrow‐derived mesenchymal stem cells

Flexor tendon injury is often associated with suboptimal outcomes and results in substantial digit dysfunction. Stem cells have been isolated from several experimental animals for the growing interest and needs of utilizing cell‐based therapies. Recently, turkey has been developed as a new large animal model for flexor tendon research. In the present study, we reported the isolation and characterization of bone marrow‐derived mesenchymal stem cells (BMSCs) from 8‐ to 12‐month‐old heritage‐breed turkeys. The isolated cells demonstrated fibroblast‐like morphology, clonogenic capacity, and high proliferation rate. These cells were positive for surface antigens CD90, CD105, and CD44, but were negative for CD45. The multipotency of turkey BMSCs was determined by differentiating cells into osteogenic, adipogenic, chondrogenic, and tenogenic lineages. There was upregulated gene expression of tenogenic markers, including mohawk, tenomodulin, and EGR1 as well as increased collagen synthesis in BMP12 induced cells. The successful isolation and verification of bone marrow‐derived MSCs from turkey would provide opportunities of studying cell‐based therapies and developing new treatments for tendon injuries using this novel preclinical large animal model. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1419–1428, 2019.     

Mechanical activation of β‐catenin regulates phenotype in adult murine marrow‐derived mesenchymal stem cells

Regulation of skeletal remodeling appears to influence the differentiation of multipotent mesenchymal stem cells (MSC) resident in the bone marrow. As murine marrow cultures are contaminated with hematopoietic cells, they are problematic for studying direct effects of mechanical input. Here we use a modified technique to isolate marrow‐derived MSC (mdMSC) from adult mice, yielding a population able to differentiate into adipogenic and osteogenic phenotypes that is devoid of hematopoietic cells. In pure mdMSC populations, a daily strain regimen inhibited adipogenic differentiation, suppressing expression of PPARγ and adiponectin. Strain increased β‐catenin and inhibition of adipogenesis required this effect. Under osteogenic conditions, strain activated β‐catenin signaling and increased expression of WISP1 and COX2. mdMSC were also generated from mice lacking caveolin‐1, a protein known to sequester β‐catenin: caveolin‐1^(?/?) mdMSC exhibited retarded differentiation along both adipogenic and osteogenic lineages but retained mechanical responses that involved β‐catenin activation. Interestingly, caveolin‐1^(?/?) mdMSC failed to express bone sialoprotein and did not form mineralized nodules. In summary, mdMSC from adult mice respond to both soluble factors and mechanical input, with mechanical activation of β‐catenin influencing phenotype. As such, these cells offer a useful model for studies of direct mechanical regulation of MSC differentiation and function. © 2010 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:1531–1538, 2010     

表皮干细胞活性复合皮肤修复裸鼠全层皮肤缺损

目的用表皮干细胞和成纤维细胞作为种子细胞，研制一种增殖能力强具有表皮、真皮的组织工程化人工复合皮肤。方法从幼儿包皮中分离表皮干细胞，用胶原Ⅳ纯化、富集表皮干细胞，接种在3T3细胞滋养层上，将体外传代培养的表皮干细胞和真皮成纤维细胞分别接种在经冷冻干燥及戊二醛交联的I型胶原基质网架的两侧，在液面下培养2周后，改为气-液界面培养，构建复合皮肤，移植到裸鼠全层皮肤缺损处，以表皮细胞胶原海绵复合皮肤和无细胞接种的胶原海绵膜作为对照。术后进行大体观察，在第7、14、21天取材行组织学、免疫组织化学及电镜观察。结果用表皮干细胞构建的复合人工皮肤增殖能力强，新生的皮肤瘢痕轻，形态满意，创面愈合的速度和质量均优于对照组。结论在胶原海绵上用表皮干细胞构建的复合人工皮肤增殖能力强，较好地解决了种子细胞的老化问题，可望成为一种较为理想的皮肤替代物。     

Transplantation KCNMA1 modified bone marrow‐mesenchymal stem cell therapy for diabetes mellitus‐induced erectile dysfunction

This study assessed the effect of KCNMA1 transfected bone marrow‐mesenchymal stem cells (BM‐MSCs) on the improvement of erectile function in diabetic rats. Sixty male Sprague–Dawley rats were injected with streptozotocin (STZ) and screened with apomorphine (APO) to establish diabetes mellitus‐induced erectile dysfunction (DMED). DMED rats were randomly divided into four groups: rats in each group underwent intracavernous injection with either phosphate buffer solution (DMED+PBS), nontransfected MSCs (DMED+MSCs), empty vector transfected MSCs (DMED+null‐MSCs) or KCNMA1 transfected MSCs (DMED+KCNMA1‐MSCs). Before injection, high levels of KCNMA1 expression were confirmed in KCNMA1‐MSCs using RT‐PCR and Western blotting. The lentivirus transfected MSCs maintained their potential for multidirectional differentiation. Four weeks after injection, erectile function was ascertained by measuring intracavernous pressure (ICP). Penile tissues were collected for immunohistochemical analysis. The expression of KCNMA1 in the corpus cavernosum was increased, and the DMED+KCNMA1‐MSCs group displayed a significant improvement of erectile function. We concluded that KCNMA1 was able to enhance the positive effect of MSCs in the treatment of diabetes‐associated erectile dysfunction.     

Acute and chronic wound fluids inversely influence adipose‐derived stem cell function: molecular insights into impaired wound healing

Wound healing is a complex biological process that requires a well‐orchestrated interaction of mediators as well as resident and infiltrating cells. In this context, mesenchymal stem cells play a crucial role as they are attracted to the wound site and influence tissue regeneration by various mechanisms. In chronic wounds, these processes are disturbed. In a comparative approach, adipose‐derived stem cells (ASC) were treated with acute and chronic wound fluids (AWF and CWF, respectively). Proliferation and migration were investigated using 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) test and transwell migration assay. Gene expression changes were analysed using quantitative real time–polymerase chain reaction. AWF had a significantly stronger chemotactic impact on ASC than CWF (77·5% versus 59·8% migrated cells). While proliferation was stimulated by AWF up to 136·3%, CWF had a negative effect on proliferation over time (80·3%). Expression of b‐FGF, vascular endothelial growth factor (VEGF) and matrix metalloproteinase‐9 was strongly induced by CWF compared with a mild induction by AWF. These results give an insight into impaired ASC function in chronic wounds. The detected effect of CWF on proliferation and migration of ASC might be one reason for an insufficient healing process in chronic wounds.     

骨髓间充质干细胞促进糖尿病溃疡创面愈合的研究现状

The delayed healing of diabetic ulcer has been haunting the surgeons and researchers for a long time.Although we have been researching and exploring the effective therapies for many years,the progress ...     

人脐带Wharton胶间质干细胞的免疫特性研究

目的 对人脐带Wharton胶间质干细胞(human umbilical cord Wharton's Jelly-derived MSC,hWJMSC)进行免疫特性的体外和体内研究,探讨其应用于临床的可行性.方法 取人脐带并分离Wharton胶,组织块法培养hWJMSC,通过细胞表面干细胞标志和多向分化实验对其进行鉴定;通过流式细胞术、免疫组织化学及RT-PCR等实验方法体外检测hWJMSC的主要组织相容性抗原(HLA-ABC和HLA-DPDQDR,即MHC-Ⅰ和MHC-Ⅱ)、表面共刺激分子(CD40、CD80、CD86)和免疫抑制分子(HLAC、IDO、PGE2)等的表达;通过细胞因子蛋白芯片和Western blot实验检测并验证hWJMSC中与免疫抑制相关的细胞因子(IL-10、TGF-β、VEGF、HGF)的表达;另外,还构建了hWJMSC-支架复合体,将其埋人家兔背部皮下,观察其在体内的免疫反应.结果 从人脐带Wharton胶中分离、培养获得长梭形细胞,经流式细胞术及多向分化实验鉴定其为间质干细胞;流式细胞术及免疫组化等检测显示所分离的hWJMSC不表达MHC-Ⅱ,表达MHC-Ⅰ,阳性表达免疫抑制因子HLA-G、IDO、PGE2以及免疫抑制相关细胞因子IL-10、TGF-β、VEGF、HGF,表明其具有低免疫原性和诱导免疫耐受的能力;hWJMSC-支架埋人家兔皮下未引起免疫反应.结论 hWJMSC的免疫原性低,具有抑制免疫排斥、诱导宿主免疫耐受的能力,可应用于异体移植.

Abstract：

Objective To probe the immunological traits of mesenchymal stem cells derived from umbilical cord Wharton's jelly (WJMSCs). Methods The diced Wharton's jelly which was from healthy fullterm birth human umbilical cord was cultured. The mesenchymal stem cells were identified with mesenchymal stem cells markers expression by flow cytometry and multiple differentiation ability. The expression of MHC- Ⅰ / Ⅱ, costimulatory molecules (CD40, CD80 and CD86) was detected with flow cytomctry, immunocytochemistry, and RT-PCR. The expression of immune inhibitors like HLA-G, IDO, and PGE2 was detected by immunocytochemistry and RT-PCR. The expression of immune-related molecules as IL-10, TGF-β, FGF and VEGF was detected with antibody microarray and western blot. Further more, to clarify the in vivo immune reaction of hWJMSCs, we fabricated the hWJMSC-scaffold constructs and implanted them into the rabbit backs. The lymphocyte infiltration and implanted cell survival observed with immunofluorescence. Results After culturinge of diced Wharton's jelly tissue, we obtained spindle-shaped cells. With differentiation medium, the cells can differentiate into osteoblasts, chongdrocytes, adipose cells and schwann cells. Expression of MHC, costimulatory molecules, and a series of immune suppressive-related molecules was found. Immune inhibitors as HLA-G, 1DO, PGE2, and immune suppressive related molecules as HGF, VEGF, TGFand IL-10 were positively expressed. But the cells did not express MHC-Ⅱ. No immune rejection was observed in vivo after implantation of hWJMSC-scaffold constructs. Conclusion It can be concluded that hWJMSCs have very low immunogenicity, which means the cells have potential to induce immune tolerance.The hWJMSCs do not provoke immune rejection in vivo.