Wang Jun-Jie, Liu Yu-Liang, Sun Yuan-Chao, Ge Wei, Wang Yong-Yong, Dyce Paul W, Hou Rong, Shen Wei
Institute of Reproductive Sciences, Qingdao Agricultural University, Qingdao, Shandong, 266109, China; Key Laboratory of Animal Reproduction and Germplasm Enhancement in Universities of Shandong, College of Animal Science and Technology, Qingdao Agricultural University, Qingdao, Shandong, 266109, China.
Chengdu Research Base of Giant Panda Breeding, Chengdu, Sichuan, 610081, China.
PLoS One. 2015 Sep 16;10(9):e0137712. doi: 10.1371/journal.pone.0137712. eCollection 2015.
It has been widely known that the giant panda (Ailuropoda melanoleuca) is one of the most endangered species in the world. An optimized platform for maintaining the proliferation of giant panda mesenchymal stem cells (MSCs) is very necessary for current giant panda protection strategies. Basic fibroblast growth factor (bFGF), a member of the FGF family, is widely considered as a growth factor and differentiation inducer within the stem cell research field. However, the role of bFGF on promoting the proliferation of MSCs derived from giant panda bone marrow (BM) has not been reported. In this study, we aimed to investigate the role of bFGF on the proliferation of BM-MSCs derived from giant panda. MSCs were cultured for cell proliferation analysis at 24, 48 and 72 hrs following the addition of bFGF. With increasing concentrations of bFGF, cell numbers gradually increased. This was further demonstrated by performing 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) cell proliferation assay, 5-Bromo-2-deoxyUridine (BrdU) labeling and cell cycle testing. Furthermore, the percentage of MSCs that were OCT4 positive increased slightly following treatment with 5 ng/ml bFGF. Moreover, we demonstrated that the extracellular signal-regulated kinase (ERK) signaling pathway may play an important role in the proliferation of panda MSCs stimulated by bFGF. In conclusion, this study suggests that giant panda BM-MSCs have a high proliferative capacity with the addition of 5 ng/ml bFGF in vitro.
众所周知，大熊猫（Ailuropoda melanoleuca）是世界上最濒危的物种之一。对于当前的大熊猫保护策略而言，一个优化的用于维持大熊猫间充质干细胞（MSCs）增殖的平台非常必要。碱性成纤维细胞生长因子（bFGF）是FGF家族的一员，在干细胞研究领域被广泛认为是一种生长因子和分化诱导剂。然而，bFGF对促进大熊猫骨髓（BM）来源的MSCs增殖的作用尚未见报道。在本研究中，我们旨在探究bFGF对大熊猫来源的BM-MSCs增殖的作用。添加bFGF后，在24、48和72小时对MSCs进行培养以进行细胞增殖分析。随着bFGF浓度的增加，细胞数量逐渐增加。通过进行3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐（MTT）细胞增殖试验、5-溴-2-脱氧尿苷（BrdU）标记和细胞周期检测进一步证实了这一点。此外，用5 ng/ml bFGF处理后，OCT4阳性的MSCs百分比略有增加。而且，我们证明细胞外信号调节激酶（ERK）信号通路可能在bFGF刺激的大熊猫MSCs增殖中起重要作用。总之，本研究表明在体外添加5 ng/ml bFGF时，大熊猫BM-MSCs具有较高的增殖能力。
