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人脐带华通氏胶干细胞在纳米纤维支架上和在序贯两阶段培养基环境中生长时,会经历增强的软骨分化。

Human umbilical cord Wharton's jelly stem cells undergo enhanced chondrogenic differentiation when grown on nanofibrous scaffolds and in a sequential two-stage culture medium environment.

机构信息

Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University of Singapore, Kent Ridge, Singapore.

出版信息

Stem Cell Rev Rep. 2012 Mar;8(1):195-209. doi: 10.1007/s12015-011-9289-8.

Abstract

The current treatments used for osteoarthritis from cartilage damage have their disadvantages of donor site morbidity, complicated surgical interventions and risks of infection and graft rejection. Recent advances in tissue engineering have offered much promise in cartilage repair but the best cell source and in vitro system have not as yet been optimised. Human bone marrow mesenchymal stem cells (hBMSCs) have thus far been the cell of choice. However, we derived a unique stem cell from the human umbilical cord Wharton's jelly (hWJSC) that has properties superior to hBMSCs in terms of ready availability, prolonged stemness characteristics in vitro, high proliferation rates, wide multipotency, non-tumorigenicity and tolerance in allogeneic transplantation. We observed enhanced cell attachment, cell proliferation and chondrogenesis of hWJSCs over hBMSCs when grown on PCL/Collagen nanoscaffolds in the presence of a two-stage sequential complex/chondrogenic medium for 21 days. Improvement of these three parameters were confirmed via inverted optics, field emission scanning electron microscopy (FESEM), MTT assay, pellet diameters, Alcian blue histology and staining, glycosaminglycans (GAG) and hyaluronic acid production and expression of key chondrogenic genes (SOX9, Collagen type II, COMP, FMOD) using immunohistochemistry and real-time polymerase chain reaction (qRT-PCR). In separate experiments we demonstrated that the 16 ng/ml of basic fibroblast growth factor (bFGF) present in the complex medium may have contributed to driving chondrogenesis. We conclude that hWJSCs are an attractive stem cell source for inducing chondrogenesis in vitro when grown on nanoscaffolds and exposed sequentially first to complex medium and then followed by chondrogenic medium.

摘要

目前用于软骨损伤性骨关节炎的治疗方法存在供体部位发病率高、手术干预复杂、感染和移植物排斥风险等缺点。组织工程学的最新进展为软骨修复带来了很大的希望,但最佳的细胞来源和体外系统尚未得到优化。人骨髓间充质干细胞(hBMSCs)是迄今为止的首选细胞。然而,我们从人脐带华通氏胶(hWJSC)中分离出一种独特的干细胞,在易于获得、体外延长干细胞特性、高增殖率、广泛多能性、非致瘤性和同种异体移植耐受性方面,其特性优于 hBMSCs。我们观察到,在存在两阶段序贯复杂/软骨形成培养基的情况下,hWJSCs 在 PCL/胶原纳米支架上培养 21 天时,其细胞黏附、细胞增殖和软骨形成能力均优于 hBMSCs。通过倒置光学显微镜、场发射扫描电子显微镜(FESEM)、MTT 测定、小球直径、阿尔辛蓝组织学和染色、糖胺聚糖(GAG)和透明质酸产生以及关键软骨形成基因(SOX9、Collagen type II、COMP、FMOD)的免疫组织化学和实时聚合酶链反应(qRT-PCR),证实了这三个参数的改善。在单独的实验中,我们证明了复杂培养基中存在的 16ng/ml 碱性成纤维细胞生长因子(bFGF)可能有助于驱动软骨形成。我们得出结论,当在纳米支架上生长并依次先暴露于复杂培养基然后再暴露于软骨形成培养基时,hWJSCs 是一种有吸引力的干细胞来源,可在体外诱导软骨形成。

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