1.Human induced pluripotent stem cells derived hepatocytes: rising promise for disease modeling, drug development and cell therapy.
Fei YI ; Guang-Hui LIU ; Juan Carlos IZPISUA BELMONTE
Protein & Cell 2012;3(4):246-250
Recent advances in the study of human hepatocytes derived from induced pluripotent stem cells (iPSC) represent new promises for liver disease study and drug discovery. Human hepatocytes or hepatocyte-like cells differentiated from iPSC recapitulate many functional properties of primary human hepatocytes and have been demonstrated as a powerful and efficient tool to model human liver metabolic diseases and facilitate drug development process. In this review, we summarize the recent progress in this field and discuss the future perspective of the application of human iPSC derived hepatocytes.
Cell Differentiation
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Cell- and Tissue-Based Therapy
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Drug Evaluation, Preclinical
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Hepatocytes
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cytology
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Humans
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Induced Pluripotent Stem Cells
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cytology
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transplantation
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Liver Diseases
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therapy
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Models, Biological
2.Direct conversion of human fibroblasts into retinal pigment epithelium-like cells by defined factors.
Kejing ZHANG ; Guang-Hui LIU ; Fei YI ; Nuria MONTSERRAT ; Tomoaki HISHIDA ; Concepcion Rodriguez ESTEBAN ; Juan Carlos IZPISUA BELMONTE
Protein & Cell 2014;5(1):48-58
The generation of functional retinal pigment epithelium (RPE) is of great therapeutic interest to the field of regenerative medicine and may provide possible cures for retinal degenerative diseases, including age-related macular degeneration (AMD). Although RPE cells can be produced from either embryonic stem cells or induced pluripotent stem cells, direct cell reprogramming driven by lineage-determining transcription factors provides an immediate route to their generation. By monitoring a human RPE specific Best1::GFP reporter, we report the conversion of human fibroblasts into RPE lineage using defined sets of transcription factors. We found that Best1::GFP positive cells formed colonies and exhibited morphological and molecular features of early stage RPE cells. Moreover, they were able to obtain pigmentation upon activation of Retinoic acid (RA) and Sonic Hedgehog (SHH) signaling pathways. Our study not only established an ideal platform to investigate the transcriptional network regulating the RPE cell fate determination, but also provided an alternative strategy to generate functional RPE cells that complement the use of pluripotent stem cells for disease modeling, drug screening, and cell therapy of retinal degeneration.
Animals
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Bestrophins
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Cell Differentiation
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Cell Line
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Cell Lineage
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Chloride Channels
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genetics
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metabolism
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Embryonic Stem Cells
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cytology
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metabolism
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Eye Proteins
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genetics
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metabolism
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Fibroblasts
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cytology
;
metabolism
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Genes, Reporter
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Green Fluorescent Proteins
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genetics
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metabolism
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Humans
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Mice
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Pigmentation
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Retinal Pigment Epithelium
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cytology
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metabolism
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Transcription Factors
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metabolism
3.Establishment of hepatic and neural differentiation platforms of Wilson's disease specific induced pluripotent stem cells.
Fei YI ; Jing QU ; Mo LI ; Keiichiro SUZUKI ; Na Young KIM ; Guang-Hui LIU ; Juan Carlos Izpisua BELMONTE
Protein & Cell 2012;3(11):855-863
The combination of disease-specific human induced pluripotent stem cells (iPSC) and directed cell differentiation offers an ideal platform for modeling and studying many inherited human diseases. Wilson's disease (WD) is a monogenic disorder of toxic copper accumulation caused by pathologic mutations of the ATP7B gene. WD affects multiple organs with primary manifestations in the liver and central nervous system (CNS). In order to better investigate the cellular pathogenesis of WD and to develop novel therapies against various WD syndromes, we sought to establish a comprehensive platform to differentiate WD patient iPSC into both hepatic and neural lineages. Here we report the generation of patient iPSC bearing a Caucasian population hotspot mutation of ATP7B. Combining with directed cell differentiation strategies, we successfully differentiated WD iPSC into hepatocyte-like cells, neural stem cells and neurons. Gene expression analysis and cDNA sequencing confirmed the expression of the mutant ATP7B gene in all differentiated cells. Hence we established a platform for studying both hepatic and neural abnormalities of WD, which may provide a new tool for tissue-specific disease modeling and drug screening in the future.
Adenosine Triphosphatases
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genetics
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metabolism
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Cation Transport Proteins
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genetics
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metabolism
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Cell Differentiation
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Copper-transporting ATPases
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Hep G2 Cells
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Hepatocytes
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cytology
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metabolism
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Hepatolenticular Degeneration
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metabolism
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pathology
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Humans
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Induced Pluripotent Stem Cells
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cytology
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Mutation
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Neural Stem Cells
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cytology
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metabolism
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Neurons
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cytology
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metabolism
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Sequence Analysis, DNA
5.Global DNA methylation and transcriptional analyses of human ESC-derived cardiomyocytes.
Ying GU ; Guang-Hui LIU ; Nongluk PLONGTHONGKUM ; Christopher BENNER ; Fei YI ; Jing QU ; Keiichiro SUZUKI ; Jiping YANG ; Weiqi ZHANG ; Mo LI ; Nuria MONTSERRAT ; Isaac CRESPO ; Antonio DEL SOL ; Concepcion Rodriguez ESTEBAN ; Kun ZHANG ; Juan Carlos IZPISUA BELMONTE
Protein & Cell 2014;5(1):59-68
With defined culture protocol, human embryonic stem cells (hESCs) are able to generate cardiomyocytes in vitro, therefore providing a great model for human heart development, and holding great potential for cardiac disease therapies. In this study, we successfully generated a highly pure population of human cardiomyocytes (hCMs) (>95% cTnT(+)) from hESC line, which enabled us to identify and characterize an hCM-specific signature, at both the gene expression and DNA methylation levels. Gene functional association network and gene-disease network analyses of these hCM-enriched genes provide new insights into the mechanisms of hCM transcriptional regulation, and stand as an informative and rich resource for investigating cardiac gene functions and disease mechanisms. Moreover, we show that cardiac-structural genes and cardiac-transcription factors have distinct epigenetic mechanisms to regulate their gene expression, providing a better understanding of how the epigenetic machinery coordinates to regulate gene expression in different cell types.
Cell Differentiation
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Cell Line
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DNA Methylation
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Embryonic Stem Cells
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cytology
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metabolism
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Epigenesis, Genetic
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Gene Expression Profiling
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Gene Expression Regulation
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Gene Regulatory Networks
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Humans
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Myocytes, Cardiac
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cytology
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metabolism
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Transcription, Genetic
6.A widely adaptable approach to generate integration-free iPSCs from non-invasively acquired human somatic cells.
Zhichao DING ; Lina SUI ; Ruotong REN ; Yanjun LIU ; Xiuling XU ; Lina FU ; Ruijun BAI ; Tingting YUAN ; Ying HAO ; Weiqi ZHANG ; Huize PAN ; Wensu LIU ; Han YU ; Concepcion Rodriguez ESTEBAN ; Xiaobing YU ; Ze YANG ; Jian LI ; Xiaomin WANG ; Juan Carlos IZPISUA BELMONTE ; Guang-Hui LIU ; Fei YI ; Jing QU
Protein & Cell 2015;6(5):386-389
Adolescent
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Adult
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Aged
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Aged, 80 and over
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Cell Culture Techniques
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methods
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Child
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Female
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Humans
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Induced Pluripotent Stem Cells
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cytology
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metabolism
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Male
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Middle Aged
7.Compound screening platform using human induced pluripotent stem cells to identify small molecules that promote chondrogenesis.
Sheng-Lian YANG ; Erica HARNISH ; Thomas LEEUW ; Uwe DIETZ ; Erika BATCHELDER ; Paul S WRIGHT ; Jane PEPPARD ; Paul AUGUST ; Cecile VOLLE-CHALLIER ; Francoise BONO ; Jean-Marc HERBERT ; Juan Carlos IZPISUA BELMONTE
Protein & Cell 2012;3(12):934-942
Articular cartilage, which is mainly composed of collagen II, enables smooth skeletal movement. Degeneration of collagen II can be caused by various events, such as injury, but degeneration especially increases over the course of normal aging. Unfortunately, the body does not fully repair itself from this type of degeneration, resulting in impaired movement. Microfracture, an articular cartilage repair surgical technique, has been commonly used in the clinic to induce the repair of tissue at damage sites. Mesenchymal stem cells (MSC) have also been used as cell therapy to repair degenerated cartilage. However, the therapeutic outcomes of all these techniques vary in different patients depending on their age, health, lesion size and the extent of damage to the cartilage. The repairing tissues either form fibrocartilage or go into a hypertrophic stage, both of which do not reproduce the equivalent functionality of endogenous hyaline cartilage. One of the reasons for this is inefficient chondrogenesis by endogenous and exogenous MSC. Drugs that promote chondrogenesis could be used to induce self-repair of damaged cartilage as a non-invasive approach alone, or combined with other techniques to greatly assist the therapeutic outcomes. The recent development of human induced pluripotent stem cell (iPSCs), which are able to self-renew and differentiate into multiple cell types, provides a potentially valuable cell resource for drug screening in a "more relevant" cell type. Here we report a screening platform using human iPSCs in a multi-well plate format to identify compounds that could promote chondrogenesis.
Cell Differentiation
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drug effects
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Chondrocytes
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cytology
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drug effects
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metabolism
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Chondrogenesis
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drug effects
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Drug Evaluation, Preclinical
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methods
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Genes, Reporter
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genetics
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Humans
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Induced Pluripotent Stem Cells
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cytology
;
drug effects
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metabolism
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Keratinocytes
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cytology
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drug effects
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metabolism
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Luciferases
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genetics
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Peptides
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chemical synthesis
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metabolism
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Reproducibility of Results
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Small Molecule Libraries
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pharmacology
8.Direct reprogramming of porcine fibroblasts to neural progenitor cells.
Xiu-Ling XU ; Ji-Ping YANG ; Li-Na FU ; Ruo-Tong REN ; Fei YI ; Keiichiro SUZUKI ; Kai LIU ; Zhi-Chao DING ; Jing QU ; Wei-Qi ZHANG ; Ying LI ; Ting-Ting YUAN ; Guo-Hong YUAN ; Li-Na SUI ; Di GUAN ; Shun-Lei DUAN ; Hui-Ze PAN ; Ping WANG ; Xi-Ping ZHU ; Nuria MONTSERRAT ; Ming LI ; Rui-Jun BAI ; Lin LIU ; Juan Carlos IZPISUA BELMONTE ; Guang-Hui LIU
Protein & Cell 2014;5(1):4-7
Animals
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Cellular Reprogramming
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Dentate Gyrus
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cytology
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Fibroblasts
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cytology
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Mice
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Neural Stem Cells
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cytology
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transplantation
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Swine
10.Modeling xeroderma pigmentosum associated neurological pathologies with patients-derived iPSCs.
Lina FU ; Xiuling XU ; Ruotong REN ; Jun WU ; Weiqi ZHANG ; Jiping YANG ; Xiaoqing REN ; Si WANG ; Yang ZHAO ; Liang SUN ; Yang YU ; Zhaoxia WANG ; Ze YANG ; Yun YUAN ; Jie QIAO ; Juan Carlos IZPISUA BELMONTE ; Jing QU ; Guang-Hui LIU
Protein & Cell 2016;7(3):210-221
Xeroderma pigmentosum (XP) is a group of genetic disorders caused by mutations of XP-associated genes, resulting in impairment of DNA repair. XP patients frequently exhibit neurological degeneration, but the underlying mechanism is unknown, in part due to lack of proper disease models. Here, we generated patient-specific induced pluripotent stem cells (iPSCs) harboring mutations in five different XP genes including XPA, XPB, XPC, XPG, and XPV. These iPSCs were further differentiated to neural cells, and their susceptibility to DNA damage stress was investigated. Mutation of XPA in either neural stem cells (NSCs) or neurons resulted in severe DNA damage repair defects, and these neural cells with mutant XPA were hyper-sensitive to DNA damage-induced apoptosis. Thus, XP-mutant neural cells represent valuable tools to clarify the molecular mechanisms of neurological abnormalities in the XP patients.
DNA Damage
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DNA Repair
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DNA-Binding Proteins
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genetics
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metabolism
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Female
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Humans
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Induced Pluripotent Stem Cells
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metabolism
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pathology
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Male
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Models, Biological
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Mutation
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Neural Stem Cells
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metabolism
;
pathology
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Xeroderma Pigmentosum
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genetics
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metabolism
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pathology