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human embryonic stem cells as an in vitro model for studying developmental origins of type 2 diabetes

World Journal of stem cells 2020年第8期12卷 761-775页

作者： Andy Chun-Hang Chen Kai Fai Lee William Shu Biu Yeung Yin Lau Lee Department of Obstetrics and Gynaecology The University of Hong KongHong KongChina Shenzhen Key Laboratory of Fertility Regulation The University of Hong Kong Shenzhen HospitalShenzhen 518053Guangdong ProvinceChina

The developmental origins of health and diseases(DOHaD)is a concept stating that adverse intrauterine environments contribute to the health risks of *** the theory emerged more than 30 years ago,many epidemiological and animal studies have confirmed that in utero exposure to environmental insults,including hyperglycemia and chemicals,increased the risk of developing noncommunicable diseases(NCDs).These NCDs include metabolic syndrome,type 2 diabetes,and complications such as diabetic *** the effects of different environmental insults on early embryo development would aid in understanding the underlying mechanisms by which these insults promote NCD *** stem cells(ESCs)have also been utilized by researchers to study the *** have pluripotent characteristics and can be differentiated into almost every cell lineage;therefore,they are excellent in vitro models for studying early developmental *** importantly,human ESCs(hESCs)are the best alternative to human embryos for research because of ethical *** this review,we will discuss different maternal conditions associated with DOHaD,focusing on the complications of maternal ***,we will review the differentiation protocols developed to generate different cell lineages from ***,we will review how hESCs are utilized as a model for research into the *** effects of environmental insults on hESC differentiation and the possible involvement of epigenetic regulation will be discussed.

关键词： Development origins of health and diseases Maternal diabetes Environmental insults Type 2 diabetes human embryonic stem cells Epigenetics

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Chemical approach to generating long-term self-renewing pMN progenitors from human embryonic stem cells

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Journal of Molecular Cell Biology 2022年第1期14卷 1-10页

作者： Guan-Yu Zhang Zhu-Man Lv Hao-Xin Ma Yu Chen Yuan Yuan Ping-Xin Sun Yu-Qi Feng Ya-Wen Li Wen-Jie Lu Yu-Dong Yang Cheng Yang Xin-Lu Yu Chao Wang Shu-Long Liang Ming-Liang Zhang Hui-Liang Li Wen-Lin Li Department of Cell Biology Second Military Medical UniversityShanghai 200433China Changhai Hospital Second Military Medical UniversityShanghai 200433China Department of Histoembryology Genetics and Developmental BiologyShanghai Jiao Tong University School of MedicineShanghai 200025China Shanghai Key Laboratory of Reproductive Medicine Shanghai Jiao Tong University School of MedicineShanghai 200025China Wolfson Institute for Biomedical Research University College LondonGower StreetLondon WC1E 6BTUK Shanghai Key Laboratory of Cell Engineering Second Military Medical UniversityShanghai 200433China

Spinal cord impairment involving motor neuron degeneration and demyelination can cause lifelong disabilities,but effective clinical interventions for restoring neurological functions have yet to be *** early spinal cord development,neural progenitors of the motor neuron(pMN)domain,defined by the expression of oligodendrocyte transcription factor 2(OLIG2),in the ventral spinal cord first generate motor neurons and then switch the fate to produce myelin-forming *** their differentiation potential,pMN progenitors could be a valuable cell source for cell therapy in relevant neurological conditions such as spinal cord ***,fast generation and expansion of pMN progenitors in vitro while conserving their differentiation potential has so far been technically *** this study,based on chemical screening,we have developed a new recipe for efficient induction of pMN progenitors from human embryonic stem *** importantly,these OLIG2+pMN progenitors can be stably maintained for multiple passages without losing their ability to produce spinal motor neurons and oligodendrocytes *** results suggest that these self-renewing pMN progenitors could potentially be useful as a renewable source of cell transplants for spinal cord injury and demyelinating disorders.

关键词： human embryonic stem cells neural differentiation chemical approach self-renewing progenitors of motor neurons oligodendrocytes

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Noggin versus basic fibroblast growth factor on the differentiation of human embryonic stem cells

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Neural Regeneration Research 2013年第23期8卷 2171-2177页

作者： Yan Zhang Junmei Zhou Zhenfu Fang Manxi Jiang Xuejin Chen Department of Laboratory Animal Sciences Shanghai Jiao Tong University School of Medicine Tissue Engineering Laboratory Shanghai Institute for Pediatric ResearchXinhua HospitalSchool of MedicineShanghai Jiao Tong University

The difference between Noggin and basic fibroblast growth factor for the neural precursor differen- tiation from human embryonic stem cells has not been studied. In this study, 100 tJg/L Noggin or 20 IJg/L basic fibroblast growth factor in serum-free neural induction medium was used to differen- tiate human embryonic stem cells H14 into neural precursors using monolayer differentiation. Two weeks after induction, significantly higher numbers of neural rosettes formed in the Noggin-induced group than the basic fibroblast growth factor-induced group, as detected by phase contrast micro- scope. Immunofluorescence staining revealed expression levels of Nestin, [3-111 Tubulin and Sox-1 were higher in the induced cells and reverse-transcription PCR showed induced cells expressed Nestin, Sox-1 and Neurofilament mRNA. Protein and mRNA expression in the Noggin-induced group was increased compared with the basic fibroblast growth factor-induced group. Noggin has a greater effect than basic fibroblast growth factor on the induction of human embryonic stem cell differentiation into neural precursors by monolayer differentiation, as Noggin accelerates and in- creases the differentiation of neural precursors.

关键词： neural regeneration stem cells basic fibroblast growth factor Noggin human embryonic stem cells neural precursors neural differentiation grants-supported paper neuroregeneration

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Consistent apparent Young’s modulus of human embryonic stem cells and derived cell types stabilized by substrate stiffness regulation promotes lineage specificity maintenance

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Cell Regeneration 2020年第1期9卷 155-170页

作者： Anqi Guo Bingjie Wang Cheng Lyu Wenjing Li Yaozu Wu Lu Zhu Ran Bi Chenyu Huang Jiao Jiao Li Yanan Du Department of Biomedical Engineering Tsinghua-Peking Center for Life SciencesMOE Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical BiologySchool of MedicineTsinghua UniversityBeijing 100084China School of Life Sciences Tsinghua UniversityBeijing 100084China Department of Biomedical Engineering McKelvey School of EngineeringWashington University in St.LouisSt.Louis 63130USA Institute of Systems Engineering Academy of Military SciencesBeijing 100071China Department of Dermatology Beijing Tsinghua Changgung HospitalSchool of Clinical MedicineTsinghua UniversityBeijing 102218China Kolling Institute University of SydneySydneyNSW 2006Australia

Background:Apparent Young’s modulus(AYM),which reflects the fundamental mechanical property of live cells measured by atomic force microscopy and is determined by substrate stiffness regulated cytoskeletal organization,has been investigated as potential indicators of cell fate in specific cell ***,applying biophysical cues,such as modulating the substrate stiffness,to regulate AYM and thereby reflect and/or control stem cell lineage specificity for downstream applications,remains a primary challenge during in vitro stem cell ***,substrate stiffness could modulate cell heterogeneity in the single-cell stage and contribute to cell fate regulation,yet the indicative link between AYM and cell fate determination during in vitro dynamic cell expansion(from single-cell stage to multi-cell stage)has not been ***:Here,we show that the AYM of cells changed dynamically during passaging and proliferation on substrates with different ***,the same change in substrate stiffness caused different patterns of AYM change in epithelial and mesenchymal cell *** stem cells and their derived progenitor cells exhibited distinguishing AYM changes in response to different substrate stiffness that had significant effects on their maintenance of pluripotency and/or lineage-specific *** substrates that were too rigid or too soft,fluctuations in AYM occurred during cell passaging and proliferation that led to a loss in lineage *** a substrate with‘optimal’stiffness(i.e.,3.5 kPa),the AYM was maintained at a constant level that was consistent with the parental cells during passaging and proliferation and led to preservation of lineage *** effects of substrate stiffness on AYM and downstream cell fate were correlated with intracellular cytoskeletal organization and nuclear/cytoplasmic localization of ***:In summary,this study suggests that optimal substrate stiffness regulated consi

关键词： Apparent Young’s modulus human embryonic stem cells Substrate stiffness YAP Cell fate

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ETV2 expression increases the efficiency of primitive endothelial cell derivation from human embryonic stem cells

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Cell Regeneration 2015年第1期4卷 1-7页

作者： Anne G Lindgren Matthew B Veldman Shuo Lin Department of Molecular Cellular and Developmental Biology University of California615 Charles E.Young Drive SouthLos AngelesCA 90095USA

Background:Endothelial cells line the luminal surface of blood vessels and form a barrier between the blood and other tissues of the *** variant 2(ETV2)is transiently expressed in both zebrafish and mice and is necessary and sufficient for vascular endothelial cell *** of this gene in early zebrafish and mouse embryos results in ectopic appearance of endothelial *** expression of ETV2 in later development results in only a subset of cells responding to the ***:We have examined the expression pattern of ETV2 in differentiating human embryonic stem cells(ESCs)to determine when the peak of ETV2 expression *** show that overexpression of ETV2 in differentiating human ESC is able to increase the number of endothelial cells generated when administered during or after the endogenous peak of gene ***:Addition of exogenous ETV2 to human ESCs significantly increased the number of cells expressing angioblast genes without arterial or venous *** may be a viable solution to generate in vitro endothelial cells for use in research and in the clinic.

关键词： human embryonic stem cells Endothelium ETV2 Differentiation

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A versatile tool for tracking the differentiation of human embryonic stem cells

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Frontiers in Biology 2010年第5期5卷 455-463页

作者： Weiqiang LI Jie QIN Xinyu LI Li ZHANG Chang LIU Fei CHEN Zifei WANG Lirong ZHANG Xiuming ZHANG Bruce TLAHN Andy Peng XIANG Center for stem Cell Biology and Tissue Engineering Sun Yat-Sen UniversityNo.74 Zhongshan Road 2Guangzhou 510080China The Key Laboratory for stem cells and Tissue Engineering Ministry of EducationGuangzhou 510080China Zhongshan Medical School Sun Yat-sen UniversityGuangzhou 510080China Department of human Genetics and Howard Hughes Medical Institute University of ChicagoChicagoIllinois 60637USA Department of Pathophysiology Guangdong College of PharmacyGuangzhou 510006China Cell Therapy Center The Third Affiliated HospitalSun Yat-sen UniversityGuangzhou 510630China

The ability of human embryonic stem cells(hESCs)to undergo indeﬁnite self-renewal in vitro and to produce lineages derived from all three embryonic germ layers both in vitro and in vivo makes such cells extremely valuable in both clinical and research ***,the generation of specialized cell lineages from a mixture of differentiated hESCs remains technically difﬁ*** speciﬁc promoter-driven reporter genes are power-ful tools for tracking cell types of interest in differentiated cell ***,we describe the construction of modular lentivectors containing different tissue-speciﬁc promoters(Tα1 ofα-tubulin;aP2 of adipocyte Protein 2;and AFP of alpha fetoprotein)driving expression of humanized Renilla greenﬂuorescent protein(hrGFP).To this end,we used MultiSite gateway technology and employed the novel vectors to successfully monitor hESC *** present a versatile method permitting target cells to be *** system will facilitate research in developmental biology,transplantation,and in vivo stem cell tracking.

关键词： human embryonic stem cells lentivector transduction greenﬂuorescent protein

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Prostaglandin E2 promotes hematopoietic development from human embryonic stem cells

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Frontiers in Biology 2010年第5期5卷 445-454页

作者： Chao YANG Jia-Fei XI Xiao-Yan XIE Wen YUE Ruo-Yong WANG Qiong WU Li-Juan HE Xue NAN Yan-Hua LI Xue-Tao PEI stem Cell and Regenerative Medicine Lab Beijing Institute of Transfusion MedicineBeijing 100850China 522 Hospital of Chinese PLA Luoyang 471000China

Recent studies have suggested that prostaglan-din(PG)E2(PGE2)and the prostaglandin pathway are essential for hematopoietic stem cell growth and ***,similar studies on hematopoietic commit-ment from human embryonic stem cells(hESCs)are still *** we report that the addition of PGE2 promotes hematopoietic differentiation of *** induced cells from hESCs/OP9 co-culture and in the presence of PGE2 were characterized by reverse transcription-PCR(RT-PCR),ﬂow cytometry,colony-forming assays and Wright-Giemsa *** results demonstrated that PGE2 exposure could alter the gene expression pattern and morphology of co-cultured hESCs and resulted in a robust hematopoietic differentiation with higher frequencies of CD34+and CD45+***,the Smad signaling pathway may be involved in PGE2 and OP9 induced hematopoietic differentiation of *** research may improve our knowledge of stem cell regulation and hopefully lead to better stem cell-based therapeutic options.

关键词： human embryonic stem cells prostaglandin E2 hematopoiesis in vitro differentiation

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Zichong granules(资冲颗粒)promote differentiation of ovarian granulosa-like cells from human embryonic stem cells in vitro

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Journal of Traditional Chinese Medicine 2021年第2期41卷 203-211页

作者： YAO Zouying WANG Jing LU Hua LI Limin ZHUO Tianxiu WAN Qian YIE Shangmian Department of Gynecology and Obstetrics The Second Medical College/Teaching HospitalChengdu University of Traditional Chinese MedicineChengdu 610041China Gynecological Department of Traditional Chinese Medicine Lishui Central HospitalLishui 323000China Department of Gynecology and Obstetrics The Affinity HospitalShaanxi University of Chinese MedicineXian 712046China Department of Gynecology and Obstetrics Affiliated HospitalChengdu University of Traditional Chinese MedicineChengdu 610072China Sichuan Academy of Traditional Chinese Medicine Chengdu 610041China

OBJECTIVE:To investigate whether Zichong granules(资冲颗粒,ZCKL),a very effective herbal formula for treating infertility,have an impact on the differentiation of ovarian granulosa cells from human embryonic stem cells(hESCs)in vitro,and to explore the cellular mechanisms of its clinical ***:Serum from ZCKL-medicated rats was prepared and used to treat mesoderm cells derived from hESCs for 6 *** rat serum and a set of growth factors were used as negative and positive controls,***:ZCKL-medicated rat serum,but not normal rat serum,induced hESCs-derived mesoderm cells to differentiate into functional ovarian granulosa-like cells(OGLCs)in a similar manner to defined growth *** induced OGLCs resembled the morphology of native human granulosa cells,expressed granulosa cell-specific markers at both the mRNA and protein levels,produced high levels of estradiol and strongly responded to follicle-stimulating hormone ***,mRNA levels of follistatin,mothers against decapentaplegic homolog 8 and bone morphogenetic protein 6 were dynamically changed during the ***:In the ZCKL treatment of infertility,one mechanism by which ZCKL may act is by influencing ovarian granulosa cell differentiation and development,possibly through the follistatin and BMP/SMAD signaling pathways.

关键词： human embryonic stem cells infertility serum mesoderm ovarian granulosa cells Zichong granules

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Effects of the Diameter and Alignment of Electrospun TSF Nanofibers on Neuronal Differentiation of human embryonic stem cells

Effects of the Diameter and Alignment of Electrospun TSF Nan...

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“细胞活动生命活力”——中国细胞生物学学会全体会员代表大会暨第十二次学术大会

作者： Rong Ye,Junxia Wang,Jing Qu,Huihui Wang,Xiaojing Xu,Huanxiang Zhang Medical College of Soochow University,Department of Cell Biology,Jiangsu Province Key Laboratory of stem Cell Research,Ren Ai Road 199,Suzhou Industrial Park,Suzhou 215123,China

While much effort has been devoted to the delineation of materials that are suitable for neuronal differentiation of human embryonic stem cells(hESCs),how material topographical features,especially the nanofiber diameter and alignment, influence the growth and neuronal differentiation of hESCs,however,is still *** the present study,we seeded hESCs-derived neural precursors on random or aligned tussah silk fibroin(TSF) electrospun nanofibers with diameters of 400 nm and 800 nm and found that the diameter and alignment of TSF scaffolds significantly influence the growth and differentiation of hESCs-derived neural *** results show that hESCs-derived neurons display good compatibility with TSF scaffolds:TSF nanofibers not only support the growth the hESCs,but guide the growth and migration of these cells and the neurite outgrowth of hESCs-derived neurons as ***,compared with cells on 800 nm aligned TSF fibers, hESCs on 400 nm aligned TSF show greater neuronal differentiation efficiency and hESCs-derived neurons on these fibers possess longer neurite ***,the property of the electrospun TSF nanofibers that can guide and direct hESC migration and differentiation,with a diameter of 400 nm in particular,makes these materials the promising candidates in nerve tissue engineering for therapeutic approaches to promote the regeneration of CNS after injury.

关键词： human embryonic stem cells TSF nanofibers Neuronal differentiation Neurite outgrowth Tissue engineering

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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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