中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (33): 5281-5285.doi: 10.3969/j.issn.2095-4344.1837

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

骨髓间充质干细胞与胰岛细胞共培养对胰岛功能的影响

杨慧明1，俞  星1，方宇辉2   

  1. 1延边大学医学院，吉林省延吉市  133002；2延边大学附属医院，吉林省延吉市  133000
  • 修回日期:2019-05-17 出版日期:2019-11-28 发布日期:2019-11-28
  • 通讯作者: 方宇辉，博士，副主任医师，延边大学附属医院，吉林省延吉市 133000
  • 作者简介:杨慧明，女，1996 年生，吉林省长春市人，汉族，延边大学医学院本科在读，主要从事自身免疫性疾病的研究。
  • 基金资助:

    吉林省教育厅项目(吉教科合字【2016】第274号)，项目负责人：方宇辉；吉林省卫生厅项目(2013Q019)，项目负责人：方宇辉；延边大学项目(延大科合字【2013】第34号)，项目负责人：方宇辉

Co-culture of bone marrow mesenchymal stem cells and islet cells influences islet function

Yang Huiming1, Yu Xing1, Fang Yuhui2   

  1. 1Yanbian University School of Medicine, Yanji 133002, Jilin Province, China; 2Affiliated Hospital of Yanbian University, Yanji 133000, Jilin Province, China
  • Revised:2019-05-17 Online:2019-11-28 Published:2019-11-28
  • Contact: Fang Yuhui, MD, Associate chief physician, Affiliated Hospital of Yanbian University, Yanji 133000, Jilin Province, China
  • About author:Yang Huiming, Yanbian University School of Medicine, Yanji 133002, Jilin Province, China
  • Supported by:

    Jilin Province Education Department Project, No. [2016]274 (to FYH); Jilin Provincial Department of Health Project, No. 2013Q019 (to FYH); Yanbian University Project, No. [2013]34 (to FYH)

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摘要：

文章快速阅读：

文题释义：
骨髓间充质干细胞：是人们在哺乳动物骨髓基质中发现的一种具有多向分化潜能的细胞亚群。它们对骨髓中的造血干细胞不仅有机械支持作用，还能分泌多种生长因子来支持造血，因而可能对受损的胰岛细胞起到滋养和保护作用。
1型糖尿病：是糖尿病的一种类型，原名胰岛素依赖型糖尿病或者青少年糖尿病。因为自体免疫系统缺陷，患者身体无法生产足够的胰岛素，导致血糖升高，容易发生酮症酸中毒，从发病开始就必须注射胰岛素，并需终身使用。

 

摘要
背景：骨髓间充质干细胞能分泌多种生长因子并支持造血，但能否保护和修复胰岛细胞功能尚不明确。
目的：探讨大鼠骨髓间充质干细胞与其胰岛细胞共培养对胰岛功能的影响。
方法：从八九周龄的雄性SD大鼠中提取股骨骨髓，采用密度梯度离心法和贴壁培养法相结合提取骨髓间充质干细胞；另取八九周龄的雄性SD大鼠，从胆总管插管注入1 g/L胶原酶P 10 mL，剪下胰腺，采用Histopaque不连续密度梯度离心法分离与纯化胰岛细胞，在显微镜下用1 mL的pipet移液器手工挑取筛选薄膜完整的胰岛细胞。将第3代骨髓间充质干细胞与胰岛细胞进行共培养，共分为4组：对照组(单纯大鼠胰岛细胞培养)、直接共培养组(接种有骨髓间充质干细胞的培养板内加入胰岛细胞进行共培养)、间接共培养组1(利用transwell培养皿进行间接共培养)、间接共培养组2(利用骨髓间充质干细胞培养液培养胰岛细胞)。培养过程中观察胰岛形态及生长状况，在培养后第3，7，14天用ELISA法检测胰岛素分泌量。实验方案经延边大学附属医院医学伦理委员会批准。
结果与结论：①4组之间采用方差分析，骨髓间充质干细胞与胰岛细胞共培养能明显提高胰岛素的分泌量，而且共培养第7天胰岛素分泌量达高峰；②在培养后第7天时，间接共培养组1的胰岛素分泌量明显高于直接共培养组，差异有显著性意义(P < 0.05)；③结果表明，骨髓间充质干细胞与胰岛细胞利用transwell培养皿进行间接共培养可以明显提高胰岛素的分泌量，而且共培养7 d效果较好。


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
ORCID: 0000-0002-3712-5240(方宇辉)

关键词: 骨髓间充质干细胞, 胰岛细胞, 直接共培养, 间接共培养, 胰岛素, SD大鼠

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells can secrete various growth factors and support hematopoiesis, but whether they can protect and repair the function of islet cells remains unclear.
OBJECTIVE: To investigate the effect of rat bone marrow mesenchymal stem cells co-cultured with islet cells on rat islet function.
METHODS: Bone marrow was extracted from male Sprague-Dawley rats aged 8-9 weeks. Bone marrow mesenchymal stem cells were isolated from bone marrow using density gradient centrifugation combined with adherent culture. Collagenase P (1 g/L, 10 mL) was injected into the common bile duct of another male Sprague-Dawley rats aged 8-9 weeks, and the pancreas was cut off to isolate and purify islet cells using Histopaque discontinuous density gradient. Islet cells with intact cell membrane were selected by hand using 1-mL pipet pipette under microscope, and co-cultured with third-generation bone marrow mesenchymal stem cells. The experiment was divided into control group (single islet cell culture), direct co-culture group (islet cells co-cultured with bone marrow mesenchymal stem cells in culture plates), indirect culture group 1 (indirect co-culture with Transwell dish), and indirect culture group 2 (islet cell culture with bone marrow mesenchymal stem cell medium). The morphology and growth of islets were observed during culture. Insulin secretion was measured by ELISA on the 3rd, 7th, and 14th days after culture. The study protocol was approved by the Ethics Committee of Yanbian University Affiliated Hospital.
RESULTS AND CONCLUSION: The results of variance analysis showed that insulin secretion was significantly increased when islet cells were co-cultured with bone marrow mesenchymal stem cells, and reached a peak on the 7th day of co-culture. Insulin secretion was significantly higher in the indirect co-culture group 1 than the direct co-culture group (P < 0.05). To conclude, the indirect co-culture method of Transwell dish can significantly improve insulin secretion, and the rational co-culture time is 7 days.

Key words: bone marrow mesenchymal stem cells, islet cells, direct co-culture, indirect co-culture, insulin, Sprague-Dawley rats

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