中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (40): 7435-7438.doi: 10.3969/j.issn.1673-8225.2010.40.006

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

CM-Dil体外标记人脐带间充质干细胞传代示踪的可行性

陈  丽，吴本清，程涵蓉，黄进洁，丁  璐，宋金枝   

  1. 暨南大学第二临床医学院，深圳市人民医院新生儿科，广东省深圳市   518020　
  • 出版日期:2010-10-01 发布日期:2010-10-01
  • 通讯作者: 吴本清，硕士，主任医师，教授，暨南大学第二临床医学院，深圳市人民医院新生儿科，广东省深圳市 518020 wubenqing783@126.com
  • 作者简介:陈丽★，女，1979年生，广东省惠州市人，汉族，暨南大学第二临床医学院在读硕士，主治医师，主要从事干细胞治疗新生鼠肺损伤方面的研究。 5823283@qq.com
  • 基金资助:

    深圳市医学重点学科专项基金(20091998 A03)；2010年深圳市科技计划项目(201002001)。

Feasibility of in vitro CM-Dil-labeled human umbilical cord mesenchymal stem cells passage tracing

Chen Li, Wu Ben-qing, Cheng Han-rong, Huang Jin-jie, Ding Lu, Song Jin-zhi   

  1. Newborn Department, Shenzhen People’s Hospital, Second College of Clinical Medicine, Jinan University, Shenzhen  518020, Guangdong Province, China
  • Online:2010-10-01 Published:2010-10-01
  • Contact: Wu Ben-qing, Master, Chief physician, Professor, Newborn Department, Shenzhen People’s Hospital, Second College of Clinical Medicine, Jinan University, Shenzhen 518020, Guangdong Province, China wubenqing783@126.com
  • About author:Chen Li★, Studying for master’s degree, Attending physician, Newborn Department, Shenzhen People’s Hospital, Second College of Clinical Medicine, Jinan University, Shenzhen 518020, Guangdong Province, China 5823283@qq.com
  • Supported by:

    the Key Special Foundation of Medical Science of Shenzhen City, No. 20091998A03*; the Science and Technology Project of Shenzhen City in 2010, No. 201002001*

PDF

453

被引次数

18

摘要：

背景：掌握人脐带间充质干细胞的移植示踪方法是研究其生物学特性的关键。
目的：观察用CM-Dil标记人脐带间充质干细胞及在体外传代示踪的可行性。
方法：采用酶消化法体外分离培养人脐带间充质干细胞，通过流式细胞仪检测细胞免疫表型和细胞周期、体外成脂成骨诱导鉴定该细胞。将第5代细胞用CM-Dil标记，并将细胞传代，荧光显微镜观察体外标记情况。
结果与结论：第3代人脐带间充质干细胞强表达CD44，CD29，低表达CD106，不表达CD34、CD40；有80%以上的细胞处在G0/G1期，成脂成骨诱导后，油红O染色和碱性磷酸酶染色分别阳性。CM-Dil标记人脐带间充质干细胞细胞标记率达90%以上，体外传代后荧光强度逐渐减退，传8代后，荧光基本消失。说明人脐带间充质干细胞增殖、分化能力强，CM-Dil标记细胞示踪方法简单易行。

关键词: 人脐带间充质干细胞, CM-Dil, 示踪, 油红O染色, 碱性磷酸酶染色

Abstract:

BACKGROUND: The key to study biological characteristics of human umbilical cord mesenchymal stem cells (hUCMSCs) is to master their tracing method of hUCMSCs transplantation.
OBJECTIVE: To analyze the tracing feasibility of CM-Dil labeled hUCMSCs in vitro.
METHODS: hUCMSCs were isolated and cultured in vitro after enzyme digestion. Immunophenotype and cell cycle were analyzed by flow cytometry, as well as induction of the adipogenic, osteogenic differentiation of hUCMSCs were identified in vitro. The fifth passage of cells were labeled with CM-Dil, passaged and observed by fluorescence microscope in vitro.  
RESULTS AND CONCLUSION: hUCMSCs at passage 3 were strongly positive for CD44 and CD29, weakly positive for CD106, but negative for CD34 and CD40. Cells in G0/G1 phase accounted for more than 80%. Following adipogenic and osteogenic induction, oil red and alkaline phosphatase staining were positive respectively. The efficiency of CM-DiI labeling hUCMSCs was above 90%. Fluorescence intensity gradually decreased in vitro. After passaged for 8 generations, it nearly quenched. These indicated that hUCMSCs have great potential to proliferation and differentiation. The tracing method of CM-DiI labeling hUCMSCs is effective and easy.

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