中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (10): 1773-1779.doi: 10.3969/j.issn.1673-8225.2012.10.015

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

人脐血间充质干细胞的生物学特性及其成骨成脂肪分化*★

张才龙1，那  娜2，张积华3，孙  康1，田少奇1，夏长所1   

  1. 青岛大学医学院附属医院，1关节外科，3健康查体中心，山东省青岛市  266003；2青岛市第八人民医院妇产科，山东省青岛市  266000
  • 收稿日期:2011-09-19 修回日期:2012-01-09 出版日期:2012-03-04 发布日期:2012-03-04
  • 通讯作者: 孙康，教授，博士生导师，青岛大学医学院附属医院关节外科，山东省青岛市 266003 并列通讯作者：田少奇，助教，青岛大学医学院附属医院关节外科，山东省青岛市 266003
  • 作者简介:张才龙★，男，1966年生，副主任医师，四川省什邡市人，汉族，1989年解放军第三军医大学毕业，硕士，主要从事关节外科及干细胞的基础与临床研究。
  • 基金资助:

    山东省科技攻关计划(2008GG30002037)，课题名称：脐血间充质干细胞移植修复骨坏死的实验研究。

Biological characteristics, osteogenic and adipogenic differentiation of mesenchymal stem cells from human umbilical cord blood*★

Zhang Cai-long1, Na Na2, Zhang Ji-hua3, Sun Kang1, Tian Shao-qi1, Xia Chang-suo1   

  1. 1Department of Joint Surgery, 3Physical Examination Center, the Affiliated Hospital of Medical College, Qingdao University, Qingdao  266003, Shandong Province, China; 2Qingdao Eighth People’s Hospital, Qingdao  266000, Shandong Province, China
  • Received:2011-09-19 Revised:2012-01-09 Online:2012-03-04 Published:2012-03-04
  • Contact: author: Sun Kang, Professor, Doctoral supervisor, Department of Joint Surgery, the Affiliated Hospital of Medical College, Qingdao University, Qingdao 266003, Shandong Province, China sunkang_qy@yahoo.com.cn author: Tian Shao-qi, Assistant teacher, Department of Joint Surgery, the Affiliated Hospital of Medical College, Qingdao University, Qingdao 266003, Shandong Province, China shaoqi99@yahoo. com.cn
  • About author:Zhang Cai-long★, Master, Associate chief physician, Department of Joint Surgery, the Affiliated Hospital of Medical College, Qingdao University, Qingdao 266003, Shandong Province, China qingyiren66@163.com
  • Supported by:

    the Key Project in Science and Technology of Shandong Province, No. 2008GG30002037*

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摘要：

背景：目前有关脐血间充质干细胞的生物学特性及分化能力的研究较少。
目的：观察人脐血间充质干细胞的生物学特性，及其向成骨、成脂肪细胞分化的能力。
方法：从不同胎龄脐血中分离间充质干细胞，对其进行原代和传代培养，并诱导其向成骨及成脂肪细胞分化。
结果与结论：倒置相差显微镜下见分离培养的脐血间充质干细胞贴壁生长，呈成纤维细胞样外观，细胞呈螺旋状排列；透射电镜下可见脐血间充质干细胞胞核比例大，细胞器少，为低分化细胞；原代及传代培养的脐血间充质干细胞生长曲线均呈S型，第3，5代细胞增殖能力最强，低胎龄的脐血间充质干细胞集落形成能力最强。流式细胞仪检测结果显示，脐血间充质干细胞稳定表达间充质干细胞相关抗原CD29，CD44 和CD90，不表达造血细胞标志CD34和CD45。成骨诱导后3 周，碱性磷酸酶染色为强阳性，茜素红染色可见大量钙化基质的形成；成脂诱导3周，油红O染色可检测到胞质中脂滴的形成。提示脐血间充质干细胞具有间充质干细胞的形态特征、生长增殖特点及细胞表面标志物等生物学特性，可向成骨细胞及脂肪细胞分化。

关键词: 脐血, 间充质干细胞, 生物学特性, 成骨细胞, 成脂肪细胞

Abstract:

BACKGROUND: Up to now, there are few reports addressing the biological properties and differentiation potential of umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs).
OBJECTIVE: To observe the biological characteristics, osteogenic and adipogenic differentiation potential of UCB-MSCs.
METHODS: MSCs were harvested and cultured from UCB at various gestational ages (GA). Harvested UCB-MSCs were cultured primarily and subcultured, and then induced to differentiate into osteoblasts and adipocytes.
RESULTS AND CONCLUSION: Under the inverted phase contrast microscope, UCB-MSCs adhered to the wall, showing fibroblast like morphology and whirlpool like growth alignment. Observation of the ultramicrostructure under transmission electron microscope showed that UCB-MSCs had a big cell nucleus, fewer cellular organelles and big karyoplasmic ratio. All of the growth curves of primary and passaged UCB-MSCs presented S-shaped. The 3rd and 5th passages of MSCs showed the strongest proliferation activity. The count of colony forming unit fibroblasts varied with GA, significant difference was found among the three GA groups (P < 0.05), and the lower GA group had a higher count of colony forming unit fibroblasts than that in the older GA group. Flow cytometry showed that these cells expressed CD29, CD44 and CD90 positively, but they failed to express hematopoiesis related molecules such as CD34 and CD45. When the MSCs were induced to osteogenic and adipogenic differentiation for 3 weeks, strong expression of alkaline phosphatase was found and the formation of a mineral extracellular matrix was detected by alizarin red staining were detected; and neutral lipid vacuoles were detected by oil red O staining. UCB-MSCs have similar morphological and biological characteristics and cell surface molecule markers with MSCs derived from bone marrow, both of which have great capability of proliferation and regeneration. UCB-MSCs can be induced to osteoblasts and adipocytes in a suitable condition in vitro.

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