Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea.
Res Vet Sci. 2012 Oct;93(2):749-57. doi: 10.1016/j.rvsc.2011.09.012. Epub 2011 Oct 8.
The present study investigated the potential of minipig bone marrow-mesenchymal stem cells (BM-MSCs) to differentiate in vitro into neuron- and cardiomyocyte-like cells. Isolated BM-MSCs exhibited a fibroblast-like morphology, expressed CD29, CD44 and CD90, and differentiated into osteocytes, adipocytes and chondrocytes. Upon induction in two different neuronal specific media, most of BM-MSCs acquired the distinctive morphological features and positively stained for nestin, neurofilament-M (NF-M), neuronal nuclei (NeuN), β-tubulin, galactocerebroside (Gal-C) and glial fibrillary acidic protein (GFAP). Expression of nestin, GFAP and NF-M was further demonstrated by RT-PCR and RT-qPCR. Following cardiomyogenic induction, MSCs exhibited a stick-like morphology with extended cytoplasmic processes, and formed cluster-like structures. The expression of cardiac specific markers α-smooth muscle actin, cardiac troponin T, desmin and α-cardiac actin was positive for immunofluorescence staining, and further confirmed by RT-PCR and RT-qPCR. In conclusion, our results showed the in vitro differentiation ability of porcine BM-MSCs into neuron-like and cardiomyocyte-like cells.
本研究探讨了小型猪骨髓间充质干细胞（BM-MSCs）在体外分化为神经元样和心肌细胞样细胞的潜力。分离的 BM-MSCs 呈成纤维细胞样形态，表达 CD29、CD44 和 CD90，并分化为成骨细胞、脂肪细胞和成软骨细胞。在两种不同的神经元特异性培养基中诱导后，大多数 BM-MSCs 获得了独特的形态特征，并对巢蛋白、神经丝-M（NF-M）、神经元核（NeuN）、β-微管蛋白、半乳糖脑苷脂（Gal-C）和神经胶质纤维酸性蛋白（GFAP）呈阳性染色。巢蛋白、GFAP 和 NF-M 的表达进一步通过 RT-PCR 和 RT-qPCR 得到证实。在心肌诱导后，MSCs 呈现出棒状形态，细胞质突起延伸，并形成簇状结构。免疫荧光染色显示心脏特异性标志物α-平滑肌肌动蛋白、心肌肌钙蛋白 T、结蛋白和α-心肌肌动蛋白的表达为阳性，并通过 RT-PCR 和 RT-qPCR 进一步证实。总之，我们的结果表明，猪 BM-MSCs 具有体外分化为神经元样和心肌细胞样细胞的能力。
