组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2014, Vol. 10 ›› Issue (6): 301-304.doi: 10.3969/j.issn.1673-0364.2014.06.02

• 论著 •    下一篇

以PECAM-1为标志去除残留未分化胚胎干细胞的研究

王玲，唐郑雅，付炜，张文杰   

  1. 上海交通大学医学院附属第九人民医院整复外科,上海市组织工程研究重点实验室；上海交通大学组织工程国家工程研究中心
  • 发布日期:2020-07-23

Removal of Ofteratoma-forming Cells from Differentiated Embryonic Stem Cells Based on PECAM-1 Expression

WANG Ling,TANG Zhengya,FU wei,ZHANG Wenjie   

  1. Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine,Shanghai Key Laboratory of Tissue Engineering,Shanghai Stem Cell Institute;Shanghai Jiaotong University,National Tissue Engineering Center of China;
  • Published:2020-07-23
  • Contact: 国家重点基础研究发展规划项目(“973”项目，2011CB964704)；国家自然科学基金(81271714，31170944)

摘要： 目的以血小板内皮细胞黏附分子-1(Platelet endothelial cell adhesion molecule-1,PECAM-1)为标志,去除小鼠胚胎干细胞(Embryonic stem cells,ESCs)中残留未分化的细胞,以去除其致瘤性,为ESCs在研究中的安全应用提供思路。方法将小鼠R1胚胎干细胞株在撤去白血病抑制因子的培养基中悬浮培养6 d,体外自发分化形成类胚体,消化打散后,以PECAM-1为标志进行磁珠分选,得到阳性与阴性细胞群体,分别以2×106个/点注射入裸鼠背部皮下,6~8周后观察畸胎瘤形成情况,组织学分析瘤体构成。结果裸鼠背部成瘤结果显示,PECAM-1+细胞群注射8个点中7个成瘤,成瘤率87.5%;而PECAM-1-细胞群注射8个点中1个成瘤,成瘤率12.5%。PECAM-1+细胞群与PECAM-1-细胞群成瘤率具有统计学差异(P=0.01)。结论应用PECAM-1可去除体外分化过程中的残留未分化ESCs,并去除其致瘤性。

关键词: 胚胎干细胞, 残留, 血小板内皮细胞黏附分子-1

Abstract: Objective To prevent teratoma formation by removal of the residual undifferentiated cells from differentiated mouse embryonic stem cells （mESCs） based on PECAM-1 expression, to hopefully promote the safe use of ESCs-based treatment in future. Methods Mouse R1 ESCs were cultured in suspension to form embryoid bodies （EBs） in the absence of leukemia inhibitory factor for 6 days. EBs were then digested into single cells and sorted by magnetic activated cell sorting （MACS） based on PECAM-1 expression. Total of 2×10^6 PECAM-1^＋ and PECAM-1^- cells were injected subcutaneously into nude mice respectively. Teratoma formation was measured after 6-8 weeks. Results Seven out of 8 injected points formed teratoma in the PECAM-1^＋ cells after 8 weeks of injection, with a tumor formation rate of 87.5%. While only 1 out of 8 injected points formed teratoma in the PECAM-1^- cells, with a tumor formation rate of 12.5%. A significantly difference was observed between PECAM-1 positive and negative cells （P=0.01）. Conclusion PECAM-1 is a specific marker for residual cells, which could be utilized to remove residual undifferentiated ESCs from in vitro differentiated ESCs, and eventually solve the tumorigenicity problem of ESCs.

Key words: Embryonic stem cells, Residual, Platelet endothelial cell adhesion molecule-1

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