Lim Ji-Hey, Boozer Lindsay, Mariani Christopher L, Piedrahita Jorge A, Olby Natasha J
North Carolina State University, College of Veterinary Medicine, Raleigh, North Carolina 27606, USA.
Cell Reprogram. 2010 Aug;12(4):417-25. doi: 10.1089/cell.2009.0093.
Adipose tissue-derived stromal cells (ADSCs) have been identified as a powerful stem cell source for cellular transplantation therapy. The dog is increasingly used as a model of human neurological disease; however, few studies have reported induction of canine ADSCs to neural lineages. We characterized canine ADSCs and investigated whether they could be induced to differentiate into neural lineages. Subcutaneous adipose tissue collected from the dorsal epaxial region of adult dogs aged from 1 to 6 years was cultured to produce ADSCs that were then induced to neural lineages. RT-PCR, flow cytometry, and immunocytochemistry were performed to characterize these cell populations. Morphologically fibroblast-like ADSCs were isolated and had similar characteristics to mesenchymal stem cells. Under neurogenic conditions containing basic fibroblast growth factor and epidermal growth factor, ADSCs formed spherical cellular aggregates that resembled neurospheres. RT-PCR confirmed expression of Sox2 and CD90 by these aggregates. Expression of neural stem/progenitor markers (Nestin, Sox2, Vimentin) and neural lineage markers (A2B5, GFAP, Tuj1) was shown on immunocytochemistry. After differentiation, 60% of the cells were Tuj1 positive. In conclusion, we isolated and generated neural progenitor cells from canine ADSCs. ADSCs have potential for future autologous cell transplantation therapy for neurological disorders.
脂肪组织来源的基质细胞（ADSCs）已被确定为细胞移植治疗的强大干细胞来源。犬越来越多地被用作人类神经疾病的模型；然而，很少有研究报道犬ADSCs向神经谱系的诱导。我们对犬ADSCs进行了表征，并研究了它们是否可以被诱导分化为神经谱系。从1至6岁成年犬的背轴上区域收集皮下脂肪组织进行培养，以产生ADSCs，然后将其诱导为神经谱系。进行逆转录聚合酶链反应（RT-PCR）、流式细胞术和免疫细胞化学以表征这些细胞群体。形态上分离出成纤维细胞样的ADSCs，其具有与间充质干细胞相似的特征。在含有碱性成纤维细胞生长因子和表皮生长因子的神经源性条件下，ADSCs形成了类似神经球的球形细胞聚集体。RT-PCR证实这些聚集体表达Sox2和CD90。免疫细胞化学显示神经干细胞/祖细胞标志物（巢蛋白、Sox2、波形蛋白）和神经谱系标志物（A2B5、胶质纤维酸性蛋白、Tuj1）的表达。分化后，60%的细胞Tuj1呈阳性。总之，我们从犬ADSCs中分离并生成了神经祖细胞。ADSCs在未来用于神经疾病的自体细胞移植治疗方面具有潜力。
