中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (10): 1737-1741.doi: 10.3969/j.issn.1673-8225.2012.10.007

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

体外诱导人骨髓间充质干细胞向Ⅱ型肺泡上皮细胞分化★

陈  寅1，2，马  南2，梅  举2，肖海波2，陆善伟1，徐怀阳1，钟  竑1，2   

  1. 1上海交通大学医学院附属新华医院(崇明)心胸外科，上海市 202150；2上海交通大学医学院附属新华医院心胸外科，上海市 200092
  • 收稿日期:2012-01-04 修回日期:2012-01-14 出版日期:2012-03-04 发布日期:2012-03-04
  • 通讯作者: 钟竑，医学博士，教授，主任医师，上海交通大学医学院附属新华医院(崇明)心胸外科，上海市 202150；上海交通大学医学院附属新华医院心胸外科，上海市 200092 zhonghongyyy@163.com
  • 作者简介:陈寅★，男，1986年生，浙江省诸暨市人，汉族，上海交通大学医学院在读硕士，主要从事心胸外科方面的研究。 zhujichenying@126.com

In vitro induction of human bone marrow mesenchymal stem cells to differentiate into type Ⅱalveolar epithelial cells

Chen Yin1, 2, Ma Nan2, Mei Ju2, Xiao Hai-bo2, Lu Shan-wei1, Xu Huai-Yang1, Zhong Hong1, 2   

  1. 1Department of Cardiothoracic Surgery, Affiliated Xinhua Hospital (Chongming Branch) of Shanghai Jiaotong University, Shanghai  202150, China; 2Department of Cardiothoracic Surgery, Affiliated Xinhua Hospital of Shanghai Jiaotong University, Shanghai  202150, China
  • Received:2012-01-04 Revised:2012-01-14 Online:2012-03-04 Published:2012-03-04
  • Contact: author: Zhong Hong, M.D., Professor, Chief physician, Department of Cardiothoracic Surgery, Affiliated Xinhua Hospital (Chongming Branch) of Shanghai Jiaotong University, Shanghai 202150, China; Department of Cardiothoracic Surgery, Affiliated Xinhua Hospital of Shanghai Jiaotong University, Shanghai 202150, China zhonghongyyy@163.com
  • About author:Chen Yin★, Studying for master’s degree, Department of Cardiothoracic Surgery, Affiliated Xinhua Hospital (Chongming Branch) of Shanghai Jiaotong University, Shanghai 202150, China; Department of Cardiothoracic Surgery, Affiliated Xinhua Hospital of Shanghai Jiaotong University, Shanghai 202150, China zhujichenying@126.com

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摘要：

背景：Ⅱ型肺泡上皮细胞被证明在大鼠肺纤维化模型中直接参与肺的修复并可以减轻肺纤维化的程度。胚胎干细胞可以在体外诱导分化Ⅱ型肺泡上皮细胞，但是其应用受到多方面的限制。
目的：探讨体外诱导骨髓间充质干细胞为Ⅱ型肺泡上皮细胞的方法及转化率。
方法：取人胸骨骨髓细胞，体外分离培养骨髓间充质干细胞。采用无血清小气道生长液和改良无血清小气道生长液作为培养液，将骨髓间充质干细胞与人胚肺间质细胞共培养。观察骨髓间充质干细胞形态并用反转录PCR检测表面活性蛋白C的mRNA以及免疫荧光检测表面活性蛋白C表达。
结果与结论：骨髓间充质干细胞与人胚肺间质细胞共培养10 d后开始出现部分骨髓间充质干细胞由原先的长梭形变成和上皮细胞相似的形态。15 d后开始检测到表面活性蛋白C mRNA，但未随共培养时间延长而有所增加。改良无血清小气道生长液与无血清小气道生长液相比能增加表面活性蛋白C mRNA表达(P < 0.05)。说明采用无血清小气道生长液或改良无血清小气道生长液，并与胚肺间质细胞共培养可以将骨髓间充质干细胞在体外诱导分化为Ⅱ型肺泡上皮细胞，但其转化率较低，表面活性蛋白C的阳性率仅为(3.15±0.69)%。

关键词: 人骨髓间充质干细胞, 共培养, 分化, Ⅱ型肺泡上皮细胞, 表面活性蛋白C

Abstract:

BACKGROUND: Alveolar epithelial type Ⅱ (AT Ⅱ) has been demonstrated to directly participate in liver recovery and can also alleviate the severity of pulmonary fibrosis in a rat model of pulmonary fibrosis. Embryonic stem cells (ESCs) can differentiate into AT Ⅱ in vitro, but the application of ESCs is confined by many factors.
OBJECTIVE: To investigate the method and transformation rate of bone marrow mesenchymalstem cells (BMSCs) differentiation into AT Ⅱ.
METHODS: The BMSCs were isolated from human bone marrow cells. BMSCs were co-cultured with human embryonic lung mesenchymal cells (MRC-5) in serum-free small airway growth medium (SAGM) and modified SAGM. The morphology of BMSCs was observed, and the expression of surfactant protein C (SPC) mRNA was detected by RT-PCR. In addition, the expression of SPC was also detected by immunofluorescence.
RESULTS AND CONCLUSION: After the BMSCs were co-cultured with MRC-5 for 10 days, some of BMSCs began to turn into epithelioid cells in morphous. After 15 days, the expression of SPC mRNA was detected, but it was not increased along with the prolongation of coculture days. Compared with SAGM, modified SAGM could significantly increase the expression of SPC mRNA (P < 0.05). BMSCs could differentiate into AT Ⅱ in SAGM or modified SAGM when co-culture with MRC-5 in vitro at low transformation rate. Positive rate of SPC was only (3.15±0.69)%.

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