中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (10): 1745-1749.doi: 10.3969/j.issn.1673-8225.2011.10.008

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

体外诱导人脐带间充质干细胞分化为许旺细胞

王广宇，赵  芳，厚荣荣，朱旅云，马利成，胡丽叶，李晓玲，杨少玲，单  巍   

  1. 白求恩国际和平医院内分泌科，河北省石家庄市 050082
  • 收稿日期:2010-10-10 修回日期:2010-11-18 出版日期:2011-03-05 发布日期:2011-03-05
  • 作者简介:王广宇★，男，1971年生，河北省廊坊市人，汉族，1998年湖北中医学院毕业，硕士，副主任医师，主要从事干细胞实验及临床研究。 yangzy_1220@yahoo.com.cn
  • 基金资助:

    河北省2007年医学科学研究重点课题计划项目(07438)。

Human umbilical cord mesenchymal stem cells differentiate into Schwann cells in vitro

Wang Guang-yu, Zhao Fang, Hou Rong-rong, Zhu Lü-yun, Ma Li-cheng, Hu Li-ye, Li Xiao-ling, Yang Shao-ling, Shan Wei   

  1. Department of Endocrinology, Bethune International Peace Hospital of Chinese PLA, Shijiazhuang  050082, Hebei Province, China
  • Received:2010-10-10 Revised:2010-11-18 Online:2011-03-05 Published:2011-03-05
  • About author:Wang Guang-yun★, Master, Associate chief physician, Department of Endocrinology, Bethune International Peace Hospital of Chinese PLA, Shijiazhuang 050082, Hebei Province, China yangzy_1220@yahoo.com.cn
  • Supported by:

    the Key Program of Medical Scientific Research of Hebei Province in 2007, No. 07438*

PDF

393

被引次数

7

摘要：

背景：人脐带Wharton’s Jelly源间充质干细胞避免了伦理的限制，来源丰富，可以作为种子细胞进行组织修复。
目的：观察体外诱导脐带Wharton’s Jelly中间充质干细胞向许旺细胞分化的可行性。 
方法：分离、培养脐带Wharton’s Jelly中间充质干细胞，流式细胞术鉴定细胞表面标志。利用神经细胞培养基、碱性成纤维生长因子、表皮生长因子、维甲酸、血小板源性生长因子等采用两步法将脐带间充质干细胞诱导分化为许旺细胞，倒置显微镜下观察细胞形态变化。利用免疫细胞化学染色法检测巢蛋白、S-100、纤维酸性蛋白的表达，反转录-聚合酶链反应、免疫印记技术检测许旺细胞特异性蛋白产物表达。
结果与结论：脐带细胞培养第7天形态发生变化，部分细胞变成梭形。原代细胞培养10 d左右可达80%~90%融合，细胞呈梭形。分离培养的细胞表达具有间充质干细胞表面特有标志：CD44(91.4%)，CD29(91.3%)，CD105(99.2%)，不表达CD34(0.2%)，CD45(0.9%)，CD14(0.6%)。脐带Wharton’s Jelly中间充质干细胞经第一阶段诱导后，细胞由短梭形变成长梭形或纺锤形，并出现聚集现象，由形状规则、表面圆滑的球形细胞团形成。第二阶段诱导后，有长梭形细胞从球形细胞团爬出，96 h后细胞形态多为长梭形，伴有多极现象。免疫细胞化学染色结果示：长梭形多极细胞具有许旺细胞特异的纤维酸性蛋白、S100蛋白染色。结果表明脐带Wharton’s Jelly中间充质干细胞可在体外诱导分化为许旺细胞。

关键词: 人脐带, Wharton&rsquo, s Jelly, 间充质干细胞, 许旺细胞, 诱导分化

Abstract:

BACKGROUND: Human umbilical cord Wharton's Jelly mesenchymal stem cells avoid the ethical constraints, which have rich resources, and can be used as seed cells for tissue repair.
OBJECTIVE: To observe the feasibility of umbilical cord Wharton's Jelly mesenchymal stem cells (UC-MSCs) differentiated into Schwann cells in vitro.
METHODS: UC-MSCs were isolated and cultivated. The surface markers of UC-MSCs were identified by flow cytometry. UC-MSCs were induced and differentiated into Schwann cells by two stages with neurons medium, basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), retinoic acid (RA), platelet-derived growth factor (PDGF). Morphologic changes of cells were observed under inverted microscope. The expression of Nestin, S-100, and glial fibrillary acidic protein (GFAP) were detected by immunocytochemical stain method. Specific proteinum expression was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot technique.
RESULTS AND CONCLUSION: Morphous of UC-MSCs were changed at 7 days culture. A part of UC-MSCs changed into fusiform shape. The primary cell could achieve 80%–90% confluence about 10 days, the morphous of UC-MSCs were fusiform shape. Cells isolated from human umbilical cord expressed specific mesenchymal stem cells surface markers: CD44 (91.4%), CD29 (91.3%), CD105 (99.2%); without expression CD34 (0.2%), CD45 (0.9%), CD14 (0.6%). Morphous of UC-MSCs changed from short fusiform shape to long fusiform shape or fusiform, cells assembled and some global cell groups formed after the first induction stage. After the second induction stage, some long fusiform shape cells grown from global cell groups. Morphous of major cells were long fusiform shape after 96 hours with the phenomenon of multipolar. The immunocytochemical stain showed that long fusiform cells have Schwann cells specific GFAP and S100 protein staining. The results showed that UC-MSCs could be differentiated into Schwann cells in vitro.

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