中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (9): 1368-1372.doi: 10.3969/j.issn.2095-4344.2017.09.011

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

腺病毒转染MafA诱导人脐带间充质干细胞表达胰腺细胞特异性基因

王鸿武1,倪  萍2,赖秀蓝1,冯学永1,林丽敏1,马  廉3,4   

  1. 1汕头大学医学院第二附属医院儿科,广东省汕头市  515041;2汕头大学医学院附属肿瘤医院乳腺肿瘤科,广东省汕头市  515031;3深圳大学妇幼保健院,广东省深圳市  518118;4深圳市坪山新区妇幼保健院,广东省深圳市  518118
  • 出版日期:2017-03-28 发布日期:2017-03-31
  • 通讯作者: 马廉,博士,主任医师,深圳大学妇幼保健院,广东省深圳市 518118;深圳市坪山新区妇幼保健院,广东省深圳市 518118
  • 作者简介:王鸿武,男,1981年生,广东省海丰县人,汉族,2009年汕头大学医学院毕业,硕士,副主任医师,主要从事小儿血液、内分泌及干细胞研究。
  • 基金资助:

    广东省科技厅社会发展计划(2013B021800256);汕头市科技计划(汕市财教〔2014〕242号);广东省医学科研基金(A2016493);汕头大学医学院临床提升计划(201410)项目

Adenovirus transfection of MafA induces human umbilical cord mesenchymal stem cells to express pancreatic cell-specific genes

Wang Hong-wu1, Ni Ping2, Lai Xiu-lan1, Feng Xue-yong1, Lin Li-min1, Ma Lian3, 4   

  1. 1Department of Pediatrics, the 2nd Affiliated Hospital of Shantou University Medical College, Shantou 515041, Guangdong Province, China; 2Department of Breast Oncology, Cancer Hospital of Shantou University Medical College, Shantou 515031, Guangdong Province, China; 3Maternity and Child Healthcare Hospital of Shenzhen University, Shenzhen 518118, Guangdong Province, China; 4Maternity and Child Healthcare Hospital of Pingshan, Shenzhen 518118, Guangdong Province, China
  • Online:2017-03-28 Published:2017-03-31
  • Contact: Ma Lian, M.D., Chief physician, Maternity and Child Healthcare Hospital of Shenzhen University, Shenzhen 518118, Guangdong Province, China; Maternity and Child Healthcare Hospital of Pingshan, Shenzhen 518118, Guangdong Province, China
  • About author:Wang Hong-wu, Master, Associate chief physician, Department of Pediatrics, the 2nd Affiliated Hospital of Shantou University Medical College, Shantou 515041, Guangdong Province, China
  • Supported by:

    the Social Development Plan of Guangdong Science and Technology Department, No. 2013B021800256; the Scientific Research Plan of Shantou City, No. (2014)242; the Medical Research Fund of Guangdong Province, No. A2016493; the Promoting Plan for Clinical Practice in Shantou University Medical College, No. 201410

摘要:

文章快速阅读:

文题释义:
腺病毒载体:
腺病毒载体转染细胞时不需要细胞因子预激,能够有效转染分裂与不分裂细胞,介导基因转移具有瞬时、高水平转基因表达的特点,腺病毒携带的转基因不整合到染色体中,所以不改变被转染细胞的基因组,因此腺病毒是基因治疗中理想的基因传递工具。
MafA基因:是一种新的胰岛β细胞特异性转录因子,只在β细胞中并通过RIPE3b1/C1调控元件来调控β细胞特异性胰岛素基因的表达,通过C1介导激活内源性胰岛素转录的主要调节剂,可促进干细胞向胰腺细胞分化。

 

摘要
背景:
人脐带间充质干细胞经化学药物或共培养等方法诱导后胰岛素分泌量低,转染胰腺发育特异性基因诱导干细胞向成熟β细胞分化是近年来的研究热点。
目的:探讨MafA基因转染后人脐带间充质干细胞向胰岛素分泌细胞分化的潜能。
方法:腺病毒携带外源性β细胞发育的关键转录因子MafA(Ad-MafA-EGFP),转染入第3代人脐带间充质干细胞,诱导后7 d观察细胞的形态学变化,应用PCR检测细胞中胰腺细胞特异性基因(glucagon、Pdx1、Nkx2.2)的表达。
结果与结论:①经Ad-MafA-EGFP转染后,倒置相差显微镜下观察人脐带间充质干细胞形态未见明显变化;②荧光显微镜下观察Ad-MafA-EGFP已进入细胞内;③经Ad-MafA-EGFP诱导后,人脐带间充质干细胞中人源性胰腺前体细胞glucagon、Pdx1、Nkx2.2基因表达增高,为人脐带间充质干细胞进一步分化成熟胰腺β细胞提供实验依据。

 

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: 0000-0002-2398-1185(王鸿武)

关键词: 干细胞, 脐带脐血干细胞, 脐带, 间充质干细胞, MafA, 胰腺前体细胞

Abstract:

BACKGROUND: Human umbilical cord mesenchymal stem cells (HUMSCs) can differentiate into insulin-producing cells after induced by chemical drugs or co-culture methods, but insulin secretion is extremely low. Therefore, to induce mature pancreatic beta cell differentiation from stem cells by adenovirus transfection of specific genes involved in the development of pancreas is a research hotspot in recent years.
OBJECTIVE: To study the differentiation potential of HUMSCs into insulin-producing cells after transfection with musculoaponeurotic fibrosarcoma oncogene homologue A (MafA).
METHODS: Ad-MafA-EGFP was transferred into passage 3 HUMSCs. After 7 days of induction, changes of cell morphology were observed by inverted phase contrast microscope. Expression of pancreatic cell-specific genes (glucagon, PDX1, Nkx2.2) was detected by PCR technique.
RESULTS AND CONCLUSION: After Ad-MafA-EGFP transfection, no significant morphological changes were observed in the HUMSCs under inverted phase contrast microscope. It was confirmed by fluorescence microscope that Ad-MafA-EGFP was transferred into the HUMSCs. After induction, the expression of human pancreatic precursor cell-related genes, including glucagon, PDX1 and Nkx2.2, was increased as detected by PCR. To conclude, these findings could provide experimental evidence for further differentiation and maturation of pancreatic β cells from HUMSCs.

 

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

Key words: Umbilical Cord, Mesenchymal Stem Cells, Maf Transcription Factors, Insulin-Secreting Cells, Tissue Engineering

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