Cruz Villagrán Claudia, Amelse Lisa, Neilsen Nancy, Dunlap John, Dhar Madhu
Department of Comparative and Experimental Medicine, University of Tennessee, Knoxville, TN 37996, USA ; Department of Large Animal Clinical Sciences, University of Tennessee, Knoxville, TN 37996, USA.
Department of Large Animal Clinical Sciences, University of Tennessee, Knoxville, TN 37996, USA.
Stem Cells Int. 2014;2014:891518. doi: 10.1155/2014/891518. Epub 2014 Nov 24.
Mesenchymal stromal cells (MSCs) are able to differentiate into extramesodermal lineages, including neurons. Positive outcomes were obtained after transplantation of neurally induced MSCs in laboratory animals after nerve injury, but this is unknown in horses. Our objectives were to test the ability of equine MSCs to differentiate into cells of neural lineage in vitro, to assess differences in morphology and lineage-specific protein expression, and to investigate if horse age and cell passage number affected the ability to achieve differentiation. Bone marrow-derived MSCs were obtained from young and adult horses. Following demonstration of stemness, MSCs were neurally induced and microscopically assessed at different time points. Results showed that commercially available nitrogen-coated tissue culture plates supported proliferation and differentiation. Morphological changes were immediate and all the cells displayed a neural crest-like cell phenotype. Expression of neural progenitor proteins, was assessed via western blot or immunofluorescence. In our study, MSCs generated from young and middle-aged horses did not show differences in their ability to undergo differentiation. The effect of cell passage number, however, is inconsistent and further experiments are needed. Ongoing work is aimed at transdifferentiating these cells into Schwann cells for transplantation into a peripheral nerve injury model in horses.
间充质基质细胞（MSCs）能够分化为包括神经元在内的外胚层谱系细胞。在神经损伤后的实验动物中移植经神经诱导的MSCs后取得了积极成果，但在马中情况尚不清楚。我们的目标是测试马MSCs在体外分化为神经谱系细胞的能力，评估形态和谱系特异性蛋白表达的差异，并研究马的年龄和细胞传代次数是否会影响分化能力。从幼年和成年马获取骨髓来源的MSCs。在证明其干性后，对MSCs进行神经诱导，并在不同时间点进行显微镜评估。结果表明，市售的氮包被组织培养板支持增殖和分化。形态学变化立即出现，所有细胞均表现出神经嵴样细胞表型。通过蛋白质印迹或免疫荧光评估神经祖细胞蛋白的表达。在我们的研究中，幼年和中年马产生的MSCs在分化能力上没有差异。然而，细胞传代次数的影响并不一致，需要进一步实验。正在进行的工作旨在将这些细胞转分化为施万细胞，以便移植到马的周围神经损伤模型中。
