羊水间充质干细胞增殖新基因Abca4及其下游调控分子在优化干细胞体外扩增与治疗方面应用的研究

申请者前期研究发现孕早期大鼠羊水间充质干细胞（AF-MSCs）增殖能力强于孕晚期，且提取成功率更高。通过对不同孕期大鼠AF-MSCs全转录组测序及生信分析，找到影响AF-MSCs增殖的基因对，并证实Abca4是新的细胞增殖相关基因，在此基础上，本课题利用转录组数据再分析，寻找Abca4调控增殖的下游效应因子，并明确它们之间的调控机制，进而对孕晚期AF-MSCs的培养条件进行优化，产生高增殖潜力的孕晚期AF-MSCs。通过本课题寻找的因子为细胞内源性存在，利用这样的差异因子对AF-MSCs培养条件进行优化避免了外源因子导入引起细胞基因组及表观遗传组的改变，提高了干细胞的安全性。如果在大鼠AF-MSCs上建系成功，进一步运用至人AF-MSCs的优化中，在既往建立的先天性脊柱裂胎鼠MSCs治疗模型上进行优化后AF-MSCs的疗效评估，最终为临床使用AF-MSCs提供一种安全的优化培养方案。

The preliminary basis of this project was the research results on my doctor's research period. I had found that the amniotic fluid mesenchymal stem/stromal cells (AF-MSCs) derived from early stage of rat pregnancy had higher proliferation than that from middle or late stages. Further research was found from the RNA sequencing that the miR-351-3p/Abca4 regulated the AF-MSCs proliferation. The Abca4 was the novel gene of proliferation associated. On the basis of above, this project re-analyzed the data from RNA sequencing, which would find out the downstream pathway of Abca4, and select the key regulator to produce the antagonistic factor of the pathway, or select the regulatory amino acid sequence to produce the peptide fragment, adding into the medium, resulting in promoting proliferation of AF-MSCs derived from the late stage of pregnancy. And further used the optimum proposal modified the human AF-MSCs, and compared the therapeutic effect, which apply a new amplification proposal for the clinical application of MSCs.