Ma Caiyun, Liu Changqing, Li Xiangchen, Lu Taofeng, Bai Chunyu, Fan Yanan, Guan Weijun, Guo Yu
Institute of Beijing Animal Science and Veterinary, Chinese Academy of Agricultural Science, Beijing, 100193, China.
Institute of Beijing Animal Science and Veterinary, Chinese Academy of Agricultural Science, Beijing, 100193, China; Department of Life Science, Department of laboratory medicine, Bengbu Medical College, Bengbu, 233030, China.
Cryobiology. 2017 Apr;75:7-14. doi: 10.1016/j.cryobiol.2017.03.003. Epub 2017 Mar 8.
Lung mesenchymal stem cells (L-MSCs) characterized by plasticity, reduced relative immune privilege and high anti-fibrosis characteristics play the crucial role in lung tissue regenerative processes. However, up to date, the multi-differentiation potentials and application values of L-MSCs are still uncertain. In the current study, the Small Tailed Han Sheep embryo L-MSCs line from 12 samples, stocking 124 cryogenically-preserved vials, was successfully established by using primary culture and cell cryopreservation techniques. Isolated L-MSCs were morphologically consistent with fibroblasts, could be passaged for at least 18 passages and more than 91.8% of cells were diploid (2n = 54) analyze by G-banding. The majority of cells were in the G0/G1 phase (70.5-91.2%), and the growth curves were all typically sigmoidal. Moreover, L-MSCs were found to express pluripotent genes Oct4, Nanog and MSCs-associated genes β-integrin, CD29, CD44, CD71, CD73 and CD90, while the expressions of hematopoietic cell markers CD34 and CD45 were negative. In addtion, the L-MSCs could be differentiated into cells of three layers with induction medium in vitro, which confirmed their multilineage differentiation potential. The secretion of urea and ALB showed the differentiated hepatocytes still possessed the detoxification function. These results indicated that the isolated L-MSCs displayed typical characteristics of mesenchymal stem cells and that the culture conditions were suitable for their maintenance of stemness and their proliferation in vitro.
肺间充质干细胞（L-MSCs）具有可塑性、相对免疫特权降低和高抗纤维化特性，在肺组织再生过程中发挥着关键作用。然而，迄今为止，L-MSCs的多向分化潜能和应用价值仍不明确。在本研究中，通过原代培养和细胞冻存技术，成功建立了来自12个样本、储存124个冻存管的小尾寒羊胚胎L-MSCs系。分离得到的L-MSCs在形态上与成纤维细胞一致，可传代至少18代，经G显带分析，超过91.8%的细胞为二倍体（2n = 54）。大多数细胞处于G0/G1期（70.5-91.2%），生长曲线均呈典型的S形。此外，发现L-MSCs表达多能基因Oct4、Nanog以及与间充质干细胞相关的基因β-整合素、CD29、CD44、CD71、CD73和CD90，而造血细胞标志物CD34和CD45的表达为阴性。此外，L-MSCs在体外诱导培养基中可分化为三层细胞，证实了它们的多系分化潜能。尿素和白蛋白的分泌表明分化后的肝细胞仍具有解毒功能。这些结果表明，分离得到的L-MSCs表现出间充质干细胞的典型特征，且培养条件适合其干性维持和体外增殖。
