Pratheesh M D, Gade Nitin E, Dubey Pawan K, Nath Amar, Sivanarayanan T B, Madhu D N, Sharma Bhaskar, Saikumar G, Sharma G Taru
Division of Physiology and Climatology, Indian Veterinary Research Institute, Izatnagar, Bareilly, U.P., India.
Vet Res Commun. 2014 Jun;38(2):139-48. doi: 10.1007/s11259-014-9597-y. Epub 2014 Mar 7.
Aim of the present study was in vitro expansion and characterization of caprine wharton's jelly derived mesenchymal stem cells (cWJ-MSCs) to investigate their tissue healing potential in xenogenic animal model. Plastic adherent fibroblastoid cell populations with distinctive homogeneous morphology were isolated from caprine Wharton's jelly explants. These Wharton's jelly derived cells were found positive for the surface markers CD-73, STRO-1 and CD-105, whereas they were negative for hematopoetic stem cell marker CD-34. In vitro cultured cWJ-MSCs also showed differentiation properties into osteogenic, adipogenic and chondrogenic lineages as demonstrated by von Kossa, Oil Red-O and Alcian blue staining respectively, which was further confirmed and quantified by flow cytometric analysis. Furthermore, these well characterized cWJ-MSCs were evaluated for the wound-healing potential in full-thickness skin wounds in rabbit model for 28 days. Caprine WJ- MSCs treated skin wounds showed significantly (P < 0.05) higher percentage of wound contraction especially at the 21(st) day post transplantation when compared to PBS treated control group animals. Further, we observed better healing potential of cWJ-MSCs in terms of histo-morphological evaluation, epithelialisation and collagenization with matured vascularization stage by day 28 as compared to control. In conclusion, cWJ- MSCs provide an alternative inexhaustible source of mesenchymal stem cells and also unravel new perspectives pertaining to the therapeutic use of these cells in different species.
本研究的目的是对山羊脐带华通氏胶间充质干细胞(cWJ-MSCs)进行体外扩增和特性鉴定,以研究其在异种动物模型中的组织修复潜力。从山羊脐带华通氏胶外植体中分离出具有独特均一形态的塑料贴壁成纤维样细胞群体。这些源自华通氏胶的细胞被发现表面标志物CD-73、STRO-1和CD-105呈阳性,而造血干细胞标志物CD-34呈阴性。体外培养的cWJ-MSCs也显示出分别向成骨、成脂和成软骨谱系分化的特性,分别通过冯科萨染色、油红O染色和阿尔辛蓝染色证明,流式细胞术分析进一步证实并量化了这一结果。此外,对这些特性明确的cWJ-MSCs在兔模型的全层皮肤伤口中的伤口愈合潜力进行了28天的评估。与磷酸盐缓冲液(PBS)处理的对照组动物相比,经山羊WJ-MSCs处理的皮肤伤口在移植后第21天显示出显著更高(P < 0.05)的伤口收缩百分比。此外,与对照组相比,我们观察到cWJ-MSCs在组织形态学评估、上皮化和胶原化方面具有更好的愈合潜力,到第28天血管化成熟。总之,cWJ-MSCs提供了一种替代的、取之不尽的间充质干细胞来源,也为这些细胞在不同物种中的治疗应用揭示了新的前景。