[1]刘卫生,王英杰,刘涛,等.模拟微重力条件下小鼠拟胚体形成及分化的初步观察[J].陆军军医大学学报(原第三军医大学学报),2008,30(17):1594-1597.
 LIU Wei-sheng,WANG Ying-jie,LIU Tao,et al.The formation and differentiation of mouse embryoid bodies under simulating microgravity[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(17):1594-1597.
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模拟微重力条件下小鼠拟胚体形成及分化的初步观察(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第17期
页码:
1594-1597
栏目:
论著
出版日期:
2008-09-15

文章信息/Info

Title:
The formation and differentiation of mouse embryoid bodies under simulating microgravity
作者:
刘卫生王英杰刘涛张世昌
第三军医大学西南医院全军感染病研究所;  延安大学附属医院心血管内科
Author(s):
LIU Wei-sheng WANG Ying-jie LIU Tao ZHANG Shi-chang
Department of Infectious Diseases, Southwest Hospital, Third Military Medical University, Chongqing 400038,Department of Cardiology, The Affiliated Hospital of Yan’an University, Yan’an  Shanxi 716000, China
关键词:
胚胎干细胞拟胚体分化生物反应器微重力
Keywords:
embryonic stem cell embryoid bodies differentiation bioreactor microgravity
分类号:
R329-33; R329.21; R339.5
文献标志码:
A
摘要:
目的     研究模拟微重力条件下小鼠胚胎干细胞(embryonic stem cell, ESc)拟胚体(embryoid bodies,EBs)的形成及分化。    方法    胰酶消化普通培养的小鼠胚胎干细胞,调整细胞数为1×106·ml-1,移入可模拟微重力的旋转型生物反应器内进行培养,显微镜动态观察鼠EBs的形成,苏木精-伊红染色观察ESc的组织细胞分化情况,检测培养过程中乳酸脱氢酶(lactate dehydrogenase,  LDH)的漏出量。取不同时相的EBs,接种6孔培养板培养3~5 d,倒置相差显微镜观察EBs来源细胞的分化。     结果    生物反应器旋转培养24 h即可肉眼观察到大量多细胞聚集体形成,相差显微镜和立体显微镜下发现,72 h后细胞聚集体进一步形成均一的EBs。随培养时间延长EBs体积不断增大,并不断有新的EBs出现。HE染色显示,EBs中的ESc呈现多组织多细胞分化特征,培养后期(12 d以后)较大EBs中心有不同程度的坏死,伴LDH漏出量的明显升高。接种EBs于6孔培养板后,镜下观察到不同类型的细胞,如内皮样细胞、成纤维样细胞、肝细胞样细胞、血管样结构以及可自发性搏动的心肌样细胞。     结论    ESc在模拟微重力条件下可大量快速形成EBs,伴有不同类型的组织细胞分化。旋转型生物反应器可作为ESc研究的有用工具。
Abstract:
Objective    To study the formation and differentiation of mouse embryoid bodies (EBs) under simulating microgravity.      Methods    Mouse embryonic stem cells (ES cells) were dissociated with trypsin and seeded onto the rotary bioreactor simulating microgravity at a concentration of 1×106·ml-1. The bioreactor was set to rotate at low speed at 37 ℃ in a humidified atmosphere containing 5% CO2. The formation and differentiation of EBs were observed under microscope and by hematoxylin/eosin (HE) staining. LDH leakage analysis was performed. The EBs were seeded in 6-well plates and cultured for 3 to 5 days. The differentiation of EBs derived cells was examined by phase contrast microscope.     Results    A lot of multicellular aggregations were seen after 24-hour rotary culture. Phase-contract microscope and stereomicroscope showed that homogeneous EBs formed from these aggregations after 72-hour culture. The volume of EBs gradually increased, accompanied with the generation of new EBs. HE staining demonstrated the characteristics of multicellular differentiation of EBs cell-derived cells. Necrosis occurred in the center of some large EBs after 12-day cultivation, meanwhile the LDH leakage was increased. Blood vessels structure, endothelioid cells, fibroblasts, hepatocyte-like cells, and spontaneously beating cardiocyte-like cells  were observed under phase-contract microscope after EBs seeding on 6-well plates.      Conclusion    Under simulating microgravity, a great quantity of EBs were differentiated from mouse ES cells and formed quickly. These results reveal that rotary bioreactor should be a useful tool for ES cell research.

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更新日期/Last Update: 2008-09-16