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

World Journal of Stem Cells 2024年第5期16卷 551-559页

作者： Amani Yehya Joseph Azar Mohamad Al-Fares Helene Boeuf Wassim Abou-Kheir Dana Zeineddine Ola Hadadeh Department of Anatomy Cell Biology and Physiological SciencesAmerican University of BeirutBeirut 1107-2020Lebanon Inserm BiotisU1026University BordeauxBordeaux F-33000France Rammal Rammal Lab Physio-Toxicity Environmental TeamFaculty of SciencesLebanese UniversityNabatieh 0000Lebanon

BACKGROUND Embryonic stem cells(ESCs)serve as a crucial ex vivo model,representing epiblast cells derived from the inner cell mass of blastocyst-stage *** exhibit a unique combination of self-renewal potency,unlimited proliferation,and *** latter is evident by the ability of the isolated cells to differ-entiate spontaneously into multiple cell lineages,representing the three primary embryonic germ *** regulatory networks guide ESCs,directing their self-renewal and lineage-specific ***,or programmed cell death,emerges as a key event involved in sculpting and forming various organs and structures ensuring proper embryonic ***-ever,the molecular mechanisms underlying the dynamic interplay between diffe-rentiation and apoptosis remain poorly *** 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,*** 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 *** hanging drop method was adopted to generate embryoid bodies(EBs)and induce terminal differentiation of *** size of the generated EBs was measured in each condition compared to the wild *** the functional level,the percentage of cardiac differentiation was measured by calculating the number of beating cardiomyocytes in the manipulated mESCs compared to the *** the molecular level,quantitative reverse transcription-polymerase chain reaction was used to assess the mRNA expression of three cardiac markers:Troponin T,GATA4,and ***,troponin T protein expression was evaluated through immunofluorescence and western blot *** Our findings showed that the upregulation of Bcl-

关键词： Mouse embryonic stem cells Self-renewal Apoptosis cardiac differentiation B-cell lymphoma 2 PIM-2 Metallothionein-1

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5-Azacytidine induces changes in electrophysiological properties of human mesenchymal stem cells

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Cell Research 2006年第12期16卷 949-960页

作者： Bartosz Balana Cecilia Nicolett Ihor Zahanich Eva M Graf Torsten Christ Sabine Boxberger Department of Pharmacology and Toxicology Medical Clinic I Medical FacultyDresden University of TechnologyFetscherstr.74D-01307 DresdenGermany

Previously,mouse bone marrow-derived stem cells（MSC）treated with the unspecific DNA methyltransferase inhibi-tor 5-azacytidine were reported to differentiate into *** aim of the present study was to investigate theefficiency of a similar differentiation strategy in human mononuclear cells obtained from healthy bone marrow *** 1-3 passages,cultures were exposed for 24 h to 5-azacytidine（3μM）followed by 6 weeks of further *** treatment did not induce expression of myogenic marker MyoD or cardiac markers Nkx2.5 and GATA-4 and didnot yield beating cells during *** patch clamp experiments,approximately 10-15% of treated and untreatedcells exhibited L-type Ca2+ *** all cells showed outwardly rectifying K+currents of rapid or slow *** current amplitude at +60 mV doubled after 6 weeks of treatment compared with time-matched *** capacitance of treated cells was significantly larger than in controls 2 weeks after treatment and remained highafter 6 *** levels ofmRNAs for the K+channels Kv1.1,Kv1.5,Kv2.1,Kv4.3 and KCNMA1 and for theCa2+ channel Cav1.2 were not affected by *** with potassium channel blockers tetraethylammoniumand clofilium at concentrations shown previously to inhibit rapid or slowly activating K+currents of hMSC inhibitedproliferation of these *** results suggest that despite the absence of differentiation of hMSC into cardiomyocytes,treatment with 5-azacytidine caused profound changes in current density.

关键词： human mesenchymal stem cells 5-azacytidine cardiac differentiation outward K~+ currents

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Developmental regulation of intracellular calcium transients during cardiomyocyte differentiation of mouse embryonic stem cells

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Acta Pharmacologica Sinica 2006年第7期27卷 901-910页

作者： Ji-dong FU Hui-mei YU Rong WANG Ji LIANG Huang-tian YANG~2 Key Laboratory of Stem Cell Biology,Institute of Health Sciences,Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences and Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China Key Laboratory of Stem Cell Biology Institute of Health Sciences Shanghai Institutes for Biological Sciences Chinese Academy of Sciences and Shanghai Jiao Tong University School of Medicine Shanghai China

Aim:To investigate the developmental regulation of intracellular Ca2+transients,an essential event in excitation-contraction coupling,during ***:Using the embryonic stem(ES)cell in vitro differentia-tion system and pharmacological intervention,we investigated the molecular andfunctional regulation of Ca2+handling proteins on the Ca2+transients at early,intermediate and later differentiation stages of ES cell-derived cardiomyocytes(ESCM).Results:Nifedipine,a selective antagonist of L-type Ca2+channels,totally blocked Ca2+transients even in the condition of field-electric stimulation inESCM at three differentiation *** Ca2+transients of ESCM were alsoinhibited by both ryanodine[an inhibitor of ryanodine receptors(RyRs)]and 2-aminoethoxydipheylborate[2-APB,an inhibitor of inositol-1,4,5-trisphosphatereceptors(IP3Rs)].The inhibitory effect of ryanodine increased with the time ofdifferentiation,while the effect of 2-APB decreased with the ***,an inhibitor of SR Ca2+-pump ATPase,inhibited Ca2+transientsequally at three differentiation stages that matched the expression ***~+free solution,which inhibits Na~+-Ca2+exchanger(NCX)to extrude Ca2+from cytosol,did not affect the amplitude of Ca2+transients of ESCM until the latter differentia-tion stage,but it significantly enhanced the basal Ca2+***:The Ca2+transients in ESCM depend on both the sarcolemmal Ca2+entry viaL-type Ca2+channels and the SR Ca2+release from RyRs and IP3Rs even at theearly differentiation stage;but NCX seems not to regulate the peak of Ca2+tran-sients until the latter differentiation stage.

关键词： Ca2+ transients cardiac differentiation embryonic stem cells Ca2+ handing proteins

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PGC-1α promotes mitochondrial respiration and biogenesis during the differentiation of hiPSCs into cardiomyocytes

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Genes & Diseases 2021年第6期8卷 891-906页

作者： Qin Zhou Hao Xu Liang Yan Liang Ye Xinyuan Zhang Bin Tan Qin Yi Jie Tian Jing Zhu Department of Pediatric Research Institute Ministry of Education Key Laboratory of Child Development and Disorders National Clinical Research Center for Child Health and Disorders(Chongqing)China International Science and Technology Cooperation Base of Child Development and Critical Disorders Children’s Hospital of Chongqing Medical UniversityChongqing400000PR China Chongqing Key Laboratory of Pediatrics Chongqing400000PR China Department of Clinical Laboratory Ministry of Education Key Laboratory of Child Development and Disorders National Clinical Research Center for Child Health and Disorders(Chongqing)China International Science and Technology Cooperation Base of Child Development and Critical Disorders Children’s Hospital of Chongqing Medical University Chongqing400000PR China Department of Cardiovascular(Internal Medicine) Ministry of Education Key Laboratory of Child Development and Disorders National Clinical Research Center for Child Health and Disorders(Chongqing)China International Science and Technology Cooperation Base of Child Development and Critical Disorders Children’s Hospital of Chongqing Medical UniversityChongqing400000PR China Chengdu Women’s and Children’s Central Hospital School of MedicineUniversity of Electronic Science and Technology of ChinaChengduSichuan Province610073PR China

Although it is widely accepted that human induced pluripotent stem cell-derived cardiomyocytes(hiPSC-CMs)are readily available,robustly reproducible,and physiologically appropriate human cells for clinical applications and research in the cardiovascular field,hiPSC-CMs cultured in vitro retain an immature metabolic phenotype that limits their application,and little is known about the underlying molecular mechanism controlling mitochondrial metabolic maturation during human induced pluripotent stem cells(hiPSCs)differentiation into *** this study,we found that peroxisome proliferator-activated receptor g coactivator-1α(PGC-1α)played an important role in inducing mitochondrial biogenesis and establishing oxidative phosphorylation(OXPHOS)during the cardiac differentiation of *** down PGC-1α by siRNA impaired mitochondrial respiration,while upregulating PGC-1α by ZLN005 promoted mitochondrial biosynthesis and function by regulating the expression of downstream genes involved in mitochondrial dynamics and oxidative metabolism in ***,we found that estrogen-related receptor a(ERRa)was required for the induction of PGC-1α stimulatory effects in *** findings provide key insights into the molecular control of mitochondrial metabolism during cardiac differentiation and may be used to generate more metabolically mature cardiomyocytes for application.

关键词： cardiac differentiation hiPSCs Mitochondrial metabolism PGC-1α Maturation

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Biomaterial property-controlled stem cell fates for cardiac regeneration

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Bioactive Materials 2016年第1期1卷 18-28页

作者： Yanyi Xu Jianjun Guan Department of Materials Science and Engineering The Ohio State UniversityColumbusOHUSA

Myocardial infarction(MI)affects more than 8 million people in the United States *** to the insufficient regeneration capacity of the native myocardium,one widely studied approach is cardiac tissue engineering,in which cells are delivered with or without biomaterials and/or regulatory factors to fully regenerate the cardiac ***,in vitro cardiac tissue engineering focuses on using biomaterials as a reservoir for cells to attach,as well as a carrier of various regulatory factors such as growth factors and peptides,providing high cell retention and a proper microenvironment for cells to migrate,grow and differentiate within the scaffolds before *** studies have shown that the full establishment of a functional cardiac tissue in vitro requires synergistic actions between the seeded cells,the tissue culture condition,and the biochemical and biophysical environment provided by the biomaterials-based *** electrical stimulation and mechanical stretch during the in vitro culture can induce the ordered orientation and differentiation of the seeded *** the other hand,the various scaffolds biochemical and biophysical properties such as polymer composition,ligand concentration,biodegradability,scaffold topography and mechanical properties can also have a significant effect on the cellular processes.

关键词： Myocardial infarction cardiac tissue engineering cardiac differentiation Biomaterials Stem cell fate

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Disruption of cardiogenesis in human embryonic stem cells exposed to trichloroethylene

Disruption of cardiogenesis in human embryonic stem cells ex...

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中国生理学会第24届全国会员代表大会暨生理学学术大会

作者：姜岩王国卿王丹苏州大学

Trichloroethylene（TCE）is ubiquitous in our living environment as a contaminant in air,water and *** exposure to TCE is reported to cause congenital heart disease in *** multiple studies have been performed in animal model test system,the mechanisms of TCE-induced cardio-teratogenicity in human remain largely unknown due to species-specific toxicological *** the current study,human embryonic stem cells（h ESCs）and cardiomyocytes（derived from the h ESCs）were used to evaluate the developmental toxicity of TCE in *** expression of genes relating to cardiac development at four different time points（day 0,day 6,day 12 and day 21）was examined tomonitor the effect of TCE on *** overall data indicated the following:（1）significant cardiac inhibition,which was characterized by decreased beating clusters and beating rates,following treatment with a non-toxic dose of TCE;（2）significant up-regulation of the Nkx2.5/Hand1 gene in cardiac progenitors and down regulation of the Mhc-7/c Tn T gene in cardiac cells;and（3）significant interference with Ca2+channel pathways in cardiomyocytes,which contributes to the adverse effect of TCE on cardiac differentiation during early embryo *** findings provide new insight into TCE biology in humans,which may help to explain the development of congenital heart defects after TCE exposure.

关键词： cardiogenesis human embryonic stem cells trichloroethylene cardiac toxicity cardiac differentiation

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