人脐带和胎盘不同层次间充质干细胞的生物学特性分析

Comparison of biological characteristics of placenta mesenchymal stem cells derived from fetus

摘要:

间充质干细胞(mesenchymal stem cell,MSC)在再生医学中具有广阔的应用前景，其临床转化应用已成为研究热点，如何大量获取原代间充质干细胞以及针对不同疾病选择最为合适的细胞来源是关键。为了探讨不同来源间充质干细胞的异同，为临床治疗与研究选择合适的种子细胞提供参考，文中比较了人脐带和胎盘不同层次间充质干细胞(MSC)的生物学特性，包括细胞形态、表面标志物、分化能力及核型分析，并对4种胎儿来源的MSCs进行了转录组序列分析，针对转录组测序结果，从增殖能力和分泌细胞因子等方面对不同来源的MSCs进行了分析验证。结果显示，脐带(umbilical cord,UC)、羊膜(amniotic membrane,AM)、绒毛膜平滑肌层(chorionic membrane,CM)、绒毛膜滋养层(chorionic villi,CV)和蜕膜(deciduae,DC)等5种不同来源的MSCs均符合2006年国际细胞治疗协会(international society for cell therapy,ISCT)的最低标准，符合干细胞的一般特性；核型分析表明除了DC来源MSCs来自母体，UC、AM、CM、CV来源MSCs均来自胎儿；转录组测序分析结果显示，来自脐带和胎盘的MSCs具有相似的基因表达模式，但也存在一些差异，这些特定基因涉及细胞周期、细胞分裂、细胞死亡、细胞生长和发育。这些基因还在转录调节、DNA修复、DNA复制和染色体稳定性中起作用，这是细胞或亚细胞组分运动、细胞通信、细胞组织突起、细胞因子分泌和激素代谢的重要组成部分。转录组测序分析的结果在一定程度上解释了不同来源的MSCs之间生物学特征的差异；基于测序结果的验证实验可知，不同来源的MSCs在增殖能力和细胞因子分泌能力存在显著差异。总的来说，UC-MSCs和CV-MSCs表现出更强的增殖能力，分泌更高水平的旁分泌因子，可能在未来疾病的治疗中有不同的优势。

Abstract:

Mesenchymal stem cells (MSCs) have broad application potentials in regenerative medicine and translational medicine.Obtaining large quantities of primary-cultured MSCs and select the most suitable cell origin for targeted diseases are critical to research.To select the most suitable seed cells of MSCs from different origins for clinical treatment and research,biological characteristics of MSCs from human umbilical cord and placenta were compared.These include cell morphology,surface marker expression,differentiation and karyotype.Transcriptome sequencing of four MSCs from fetus were performed and the results were analyzed from the perspective of proliferation and cytokine secretion.The results revealed that MSCs from umbilical cord (UC),amniotic membrane (AM),chorionic membrane (CM),chorionic villi (CV) and deciduae (DC) met the minimum standards of the International Society of Cell Therapy (ISCT) in 2006 and had the general characteristics of stem cells.Karyotype analysis showed that MSCs derived from UC,AM,CM and CV were all from fetus except that the DC-MSCs were from mother.Transcriptome sequencing analysis showed that hMSCs from umbilical cord and placenta had similar gene expression patterns,while different expression patterns were observed in specific genes involved in cell cycle,cell division,cell death,cell growth and development.These genes play important roles in transcriptional regulation,DNA repair,DNA replication and chromosome stability,which were momentous components of cellular or subcellular fraction movement,cell communication,cell tissue protrusions,cytokine secretion and hormone metabolism.Transcriptome sequencing analysis explained the differences in biological characteristics among MSCs from different sources,while verification experiments based on the transcriptome sequencing results showed that the proliferation and cytokine secretion capabilities of MSCs from different sources were significantly different.In all,UC-MSCs and CV-MSCs with stronger proliferation and higher levels of paracrine factors secretion may show their respective advantages in treating diseases.