中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (7): 1032-1037.doi: 10.12307/2022.140

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

植物源性碱性成纤维细胞生长因子维持人胚胎干细胞的生长与分化

刘  峰1,2,彭宇环3,罗良平1,吴本清4   

  1. 1暨南大学附属第一医院医学影像中心,广东省广州市  510630;中国科学院大学深圳医院(光明),2内分泌科,3药学部,4新生儿科,广东省深圳市  518106
  • 收稿日期:2020-10-19 修回日期:2020-10-22 接受日期:2020-11-21 出版日期:2022-03-08 发布日期:2021-10-29
  • 通讯作者: 吴本清,博士,主任医师,中国科学院大学深圳医院(光明)新生儿科,广东省深圳市 518106
  • 作者简介:刘峰,男,1980年生,江西省吉安市人,2010年中南大学毕业,博士,主要从事内分泌代谢的基础与临床研究。

Plant-derived basic fibroblast growth factor maintains the growth and differentiation of human embryonic stem cells

Liu Feng1, 2, Peng Yuhuan3, Luo Liangping1, Wu Benqing4   

  1. 1Department of Medical Imaging Center, First Affiliated Hospital, Jinan University, Guangzhou 510630, Guangdong Province, China; 2Department of Endocrinology, 3Department of Pharmacy, 4Department of Neonatology, Shenzhen Hospital, University of Chinese Academy of Sciences, Shenzhen 518106, Guangdong Province, China
  • Received:2020-10-19 Revised:2020-10-22 Accepted:2020-11-21 Online:2022-03-08 Published:2021-10-29
  • Contact: Wu Benqing, MD, Chief physician, Department of Neonatology, Shenzhen Hospital, University of Chinese Academy of Sciences, Shenzhen 518106, Guangdong Province, China
  • About author:Liu Feng, MD, Department of Medical Imaging Center, First Affiliated Hospital, Jinan University, Guangzhou 510630, Guangdong Province, China; Department of Endocrinology, Shenzhen Hospital, University of Chinese Academy of Sciences, Shenzhen 518106, Guangdong Province, China

摘要:

文题释义:
碱性成纤维细胞生长因子:1974年GOSPODAROWICZ首次从牛脑垂体的组织提取液中分离获得碱性成纤维细胞生长因子,它是成纤维细胞生长因子家族的一个成员,定位在4号染色体的短臂上,由155个氨基酸组成。碱性成纤维细胞生长因子是重要的促有丝分裂因子,具备促进血管和组织再生的生物学功能。人胚胎干细胞的常规培养体系需要4 μg/L碱性成纤维细胞生长因子和小鼠胚胎成纤维细胞作为支撑,来维持其未分化状态,碱性成纤维细胞生长因子信号对维持人胚胎干细胞的自我更新起关键作用。
人胚胎干细胞自我更新和分化:1998年THOMSON建立了首株人类胚胎干细胞系,它的显著特征是具有自我更新能力和多向分化潜能。自我更新指干细胞通过对称或者不对称分裂产生至少一个保留干细胞特性子细胞的过程,在人胚胎干细胞中,通过Oct4、Sox2和Nanog这些转录因子所调控的基因能够在维持干细胞干性的同时抑制促进分化基因的表达,实现人胚胎干细胞在体外长期传代,并具备向内、中、外3个胚层分化的能力。
背景:碱性成纤维细胞生长因子是维持人胚胎干细胞长期培养的重要生长因子,提供来源可靠、无动物源性以及低成本的碱性成纤维细胞生长因子产品对维持人胚胎干细胞生长以及规模化制备至关重要。
目的:为了排除动物源性或者其他细菌类的潜在干扰,该研究评估植物源性碱性成纤维细胞生长因子对人胚胎干细胞生长与分化的影响。
方法:以人胚胎干细胞系(H9)作为研究材料,采用免疫荧光染色、流式细胞仪检测、碱性磷酸酶染色、RT-PCR方法比较不同质量浓度植物源性碱性成纤维细胞生长因子维持干细胞特性的能力以及其向外、中、内三胚层分化和胰腺细胞分化的潜能。
结果与结论:植物源性碱性成纤维细胞生长因子(4 μg/L和0.4 μg/L)和常规碱性成纤维细胞生长因子(4 μg/L)都能很好地维持人胚胎干细胞的生长和分化能力,提示植物源性碱性成纤维细胞生长因子可以作为有效支持人胚胎干细胞生长和分化的备选产品。
https://orcid.org/0000-0002-8543-6078(刘峰) 

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

关键词: 干细胞, 人胚胎干细胞, 碱性成纤维细胞生长因子, 植物源性, 分化, 胰腺细胞

Abstract: BACKGROUND: Basic fibroblast growth factor is an important growth factor to maintain the long-term culture of human embryonic stem cells. Providing reliable, non-animal-derived and low-cost basic fibroblast growth factor products is essential for maintaining the growth of human embryonic stem cells and large-scale production.
OBJECTIVE: To eliminate the potential interference of animal or other bacteria, this study mainly evaluated the effects of plant-derived basic fibroblast growth factor on the growth and differentiation of human embryonic stem cells. 
METHODS: Human embryonic stem cells line (H9) was used as research materials. Immunofluorescence staining, flow cytometry detection, alkaline phosphatase staining, and RT-PCR were utilized to systematically compare the ability of plant-derived basic fibroblast growth factor to maintain the characteristics of stem cells at different concentrations and its potential to differentiate into ectoderm, mesoderm, endoderm and pancreatic cells. 
RESULTS AND CONCLUSION: Plant-derived basic fibroblast growth factor (4 μg/L and 0.4 μg/L) and basic fibroblast growth factor (4 μg/L) used in conventional culture system could maintain the growth and differentiation of human embryonic  stem cells, indicating that plant-derived basic fibroblast growth factor can be used as an effective alternative product to support the growth and differentiation of human embryonic stem cells. 

Key words: stem cells, human embryonic stem cells, basic fibroblast growth factor, plant-derived, differentiation, pancreatic cells

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