Liu Xiaoyu, Zhang Tao, Wang Rui, Shi Ping, Pan Bochen, Pang Xining
Reproductive medicine Center, Obstetrics and Gynecology Department, Shengjing Hospital affiliated to China Medical University, China.
Department of Stem Cells and Regenerative Medicine, Key Laboratory of Cell Biology, Chinese Ministry of Public Health and Key Laboratory of Medical Cell Biology, Chinese Ministry of Education, China Medical University, China.
Ann Clin Lab Sci. 2019 Jan;49(1):63-71.
This study aimed to evaluate the use of Insulin-Transferrin-Selenium (ITS) medium in place of fetal bovine serum (FBS) to culture human amnion mesenchymal stem cells (hAMSCs). Cell morphology, ultrastructure, proliferation, migration and MSC related markers were assessed accordingly. The hAMSCs were induced to osteocyte, chondrocyte, adipocyte and keratinocyte by culturing in appropriate induction medium. hAMSCs mRNA expression was detected for the matrix metalloproteinases 2 (MMP2), keratinocyte growth factor (KGF), vascular endothelial growth factor (VEGF), insulin-like growth factor-I (IGF-I), Platelet-derived Growth Factor (PDGF), and transforming growth factor beta 1 (TGF-β) by real-time quantitative RT-PCR. Our results showed that hAMSCs cultured in ITS medium exhibited similar proliferation rates, demonstrated a statistically significant increased migration and expressed similar levels of MSC markers(CD73+, CD90+, CD105+, CD45-, CD34-) compared with those cultured in FBS. Osteoblasts, chondrocytes, adipocytes and keratinocytes were differentiated. Results of transmission electron microscope (TEM) revealed that hAMSCs cultured in ITS medium underwent active metabolism. The mRNA expression of MMP2, VEGF, KGF, TGF-β, IGF-I and PDGF upregulated in ITS medium. In conclusion, ITS medium has the potential to be used for the expansion of hAMSCs before clinical application.
本研究旨在评估使用胰岛素-转铁蛋白-硒（ITS）培养基替代胎牛血清（FBS）培养人羊膜间充质干细胞（hAMSCs）的效果。据此评估细胞形态、超微结构、增殖、迁移及间充质干细胞相关标志物。通过在适当的诱导培养基中培养，将hAMSCs诱导分化为骨细胞、软骨细胞、脂肪细胞和角质形成细胞。采用实时定量逆转录聚合酶链反应（RT-PCR）检测hAMSCs中基质金属蛋白酶2（MMP2）、角质形成细胞生长因子（KGF）、血管内皮生长因子（VEGF）、胰岛素样生长因子-I（IGF-I）、血小板衍生生长因子（PDGF）和转化生长因子β1（TGF-β）的mRNA表达。我们的结果显示，与在FBS中培养的hAMSCs相比，在ITS培养基中培养的hAMSCs具有相似的增殖率，迁移能力显著增强，且MSC标志物（CD7 +、CD90 +、CD105 +、CD45 -、CD34 -）表达水平相似。成功分化出了成骨细胞、软骨细胞、脂肪细胞和角质形成细胞。透射电子显微镜（TEM）结果显示，在ITS培养基中培养的hAMSCs代谢活跃。ITS培养基中MMP2、VEGF、KGF、TGF-β、IGF-I和PDGF的mRNA表达上调。总之，ITS培养基在临床应用前有潜力用于hAMSCs的扩增。
