Amniotic fluid mesenchymal stromal cells from early stages of embryonic development have higher self-renewal potential
Amniotic fluid mesenchymal stromal cells from early stages of embryonic development have higher self-renewal potential
复制标题
来自胚胎发育早期阶段的羊水间充质基质细胞具有更高的自我更新潜力
DOI：
10.1007/s11626-020-00511-z
复制
发表时间：
2020-10-07
影响因子：
2.1
通讯作者：
Yuan, Zhengwei
中科院分区：
文献类型：
--
作者：
Huang, Jieting;Ma, Wei;Yuan, Zhengwei
羊水(AF)是间充质基质细胞(MSCs)的丰富来源，具有向多个谱系分化的能力，使其成为一种有前景的强大再生医学工具。然而，关于来自不同妊娠阶段的AFMSCs之间的差异的信息有限。本研究分离并培养了125只胎龄为12天、15天、18天和21天的大鼠胚胎的AFMSCs。原代E15细胞体积最小，最容易培养，通常呈球形生长，类似于胚胎干细胞(ESCs)的生长形态。E12和E15 AFMSCs贴壁后生长速度较快，可传代60次以上，仍保持连续增殖状态，而E18和E21来源的AFMSCs一般只能维持10代。为了确定这种差异的可能原因，我们使用RT-qPCR检测了几个与自我更新能力和细胞起源相关的基因。SOX2的表达水平表明E12和E15的AFMSCs具有较强的自我更新能力。K19、Col2A1、FGF5、AFP和SPC的表达水平表明AFMSC培养中存在混合群体细胞共存。总之，E15细胞比E12细胞更容易培养，传代次数更多，Sox2的表达也比E18或E21细胞高。E15来源的AFMSCs较晚期来源的细胞具有更高的存活率和增殖能力。因此，早期的房颤细胞可能是探索涉及AFMSCs的潜在治疗方法的良好选择。
Amniotic fluid (AF) is a rich source of mesenchymal stromal cells (MSCs) that have the ability to differentiate into multiple lineages rendering them a promising and powerful tool for regenerative medicine. However, information regarding the differences among AFMSCs derived from different gestational stages is limited. In the present study, AFMSCs derived from 125 pregnant rats at four embryonic day (E) stages (E12, E15, E18, and E21) were isolated and cultured. The primary E15 cells were the smallest in size and the easiest to culture and usually grew in a spherical shape that resembled the growth morphology of embryonic stem cells (ESCs). Once adhered, the E12 and E15 AFMSCs grew faster and could be passaged more than 60 times while still maintaining a continuous proliferative state; however, AFMSCs derived from E18 and E21 could normally be maintained for only 10 passages. To identify the possible reasons for this difference, RT-qPCR was used to examine several genes associated with self-renewal ability and cell origin. The Sox2 expression levels indicated that AFMSCs from E12 and E15 possessed stronger self-renewal capability. The K19, Col2A1, FGF5, AFP, and SPC expression levels indicated there were mixed-population cells co-existing in the AFMSC culture. In conclusion, E15 cells were easier to culture than E12 cells, could be passaged more often, and had a higher Sox2 expression than E18 or E21 cells. The E15-derived AFMSCs had higher viability and proliferative capacity than cells from the later stages. Therefore, AF cells from the early stages could be a good choice for exploring potential treatments involving AFMSCs.