Cardiac differentiation is modulated by anti-apoptotic signals in murine embryonic stem cells  

作　　者：Amani Yehya Joseph Azar Mohamad Al-Fares Helene Boeuf Wassim Abou-Kheir Dana Zeineddine Ola Hadadeh 

机构地区：[1]Department of Anatomy,Cell Biology and Physiological Sciences,American University of Beirut,Beirut 1107-2020,Lebanon [2]Inserm,Biotis,U1026,University Bordeaux,Bordeaux F-33000,France [3]Rammal Rammal Lab,Physio-Toxicity Environmental Team,Faculty of Sciences,Lebanese University,Nabatieh 0000,Lebanon

出　　处：《World Journal of Stem Cells》2024年第5期551-559,共9页世界干细胞杂志（英文版）（电子版）

基　　金：Supported by the National Council for Scientific Research in Lebanon,CNRS-L.

摘　　要：BACKGROUND Embryonic stem cells(ESCs)serve as a crucial ex vivo model,representing epiblast cells derived from the inner cell mass of blastocyst-stage embryos.ESCs exhibit a unique combination of self-renewal potency,unlimited proliferation,and pluripotency.The latter is evident by the ability of the isolated cells to differ-entiate spontaneously into multiple cell lineages,representing the three primary embryonic germ layers.Multiple regulatory networks guide ESCs,directing their self-renewal and lineage-specific differentiation.Apoptosis,or programmed cell death,emerges as a key event involved in sculpting and forming various organs and structures ensuring proper embryonic development.How-ever,the molecular mechanisms underlying the dynamic interplay between diffe-rentiation and apoptosis remain poorly understood.AIM To investigate the regulatory impact of apoptosis on the early differentiation of ESCs into cardiac cells,using mouse ESC(mESC)models-mESC-B-cell lym-phoma 2(BCL-2),mESC-PIM-2,and mESC-metallothionein-1(MET-1)-which overexpress the anti-apoptotic genes Bcl-2,Pim-2,and Met-1,respectively.METHODS mESC-T2(wild-type),mESC-BCL-2,mESC-PIM-2,and mESC-MET-1 have been used to assess the effect of potentiated apoptotic signals on cardiac differentiation.The hanging drop method was adopted to generate embryoid bodies(EBs)and induce terminal differentiation of mESCs.The size of the generated EBs was measured in each condition compared to the wild type.At the functional level,the percentage of cardiac differentiation was measured by calculating the number of beating cardiomyocytes in the manipulated mESCs compared to the control.At the molecular level,quantitative reverse transcription-polymerase chain reaction was used to assess the mRNA expression of three cardiac markers:Troponin T,GATA4,and NKX2.5.Additionally,troponin T protein expression was evaluated through immunofluorescence and western blot assays.RESULTS Our findings showed that the upregulation of Bcl-2,Pim-2,and Met-1 genes led to a reduction in th

关 键 词：Mouse embryonic stem cells SELF-RENEWAL Apoptosis Cardiac differentiation B-cell lymphoma 2 PIM-2 Metallothionein-1 

分 类 号：R730.5[医药卫生—肿瘤]