Toupadakis Chrisoula A, Wong Alice, Genetos Damian C, Cheung Whitney K, Borjesson Dori L, Ferraro Gregory L, Galuppo Lawrence D, Leach J Kent, Owens Sean D, Yellowley Clare E
Department of Anatomy, University of California-Davis, Davis, CA 95616, USA.
Am J Vet Res. 2010 Oct;71(10):1237-45. doi: 10.2460/ajvr.71.10.1237.
To determine the optimal osteogenic source of equine mesenchymal stem cells (eMSCs) and optimize collection of and expansion conditions for those cells.
10 adult Quarter Horses and 8 newborn Thoroughbred foals.
eMSCs were isolated from bone marrow (BM), adipose tissue, and umbilical cord blood and tissue, and the osteogenic potential of each type was assessed. Effects of anatomic site, aspiration volume, and serum type on eMSC yield from BM were investigated.
BM-eMSCs had the highest overall expression of the osteogenic genes Cbfa1, Osx, and Omd and staining for ALP activity and calcium deposition. There was no significant difference in BM-eMSC yield from the tuber coxae or sternum, but yield was significantly greater from the first 60-mL aspirate than from subsequent aspirates. The BM-eMSC expansion rate was significantly higher when cells were cultured in fetal bovine serum instead of autologous serum (AS).
eMSCs from BM possessed the highest in vitro osteogenic potential; eMSCs from adipose tissue also had robust osteogenic potential. The tuber coxae and the sternum were viable sources of BM-eMSCs in yearlings, and 60 mL of BM aspirate was sufficient for culture and expansion. Expanding BM-eMSCs in AS to avoid potential immunologic reactions decreased the total yield because BM-eMSCs grew significantly slower in AS than in fetal bovine serum. Additional studies are needed to determine optimal ex vivo eMSC culture and expansion conditions, including the timing and use of growth factor—supplemented AS.
确定马间充质干细胞（eMSCs）的最佳成骨来源，并优化这些细胞的采集和扩增条件。
10匹成年四分之一马和8匹新生纯种马驹。
从骨髓（BM）、脂肪组织以及脐带血和脐带组织中分离eMSCs，并评估每种类型的成骨潜力。研究了解剖部位、抽吸量和血清类型对BM中eMSC产量的影响。
BM-eMSCs在成骨基因Cbfa1、Osx和Omd的总体表达以及碱性磷酸酶活性和钙沉积染色方面表现最高。来自髋结节或胸骨的BM-eMSC产量无显著差异，但首次60 mL抽吸物的产量显著高于后续抽吸物。当细胞在胎牛血清而非自体血清（AS）中培养时，BM-eMSC的扩增率显著更高。
来自BM的eMSCs具有最高的体外成骨潜力；来自脂肪组织的eMSCs也具有强大的成骨潜力。髋结节和胸骨是一岁马中BM-eMSCs的可行来源，60 mL BM抽吸物足以用于培养和扩增。在AS中扩增BM-eMSCs以避免潜在的免疫反应会降低总产量，因为BM-eMSCs在AS中的生长明显慢于胎牛血清中的生长。需要进一步研究以确定最佳的体外eMSC培养和扩增条件，包括补充生长因子的AS的使用时机。
