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Combination of cell signaling molecules can facilitate MYOD1-mediated myogenic transdifferentiation of pig fibroblasts

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Journal of Animal Science and Biotechnology 2021年第4期12卷 1662-1674页

作者： Jinsol Jeong Kwang-Hwan Choi Seung-Hun Kim Dong-Kyung Lee Jong-Nam Oh Mingyun Lee Gyung Cheol Choe Chang-Kyu Lee Department of Agricultural Biotechnology Animal Biotechnology Majorand Research Institute of Agriculture and Life ScienceSeoul National UniversitySeoul 08826South Korea Present address:Research and Development Center Space F corporationHwasung-siGyeonggi-do 18471South Korea Institute of Green Bio Science and Technology Seoul National UniversityPyeong ChangKangwon-do 25354South Korea

Background:Myogenic transdifferentiation can be accomplished through ectopic MYOD1 expression,which is facilitated by various signaling pathways associated with *** this study,we attempted to transdifferentiate pig embryonic fibroblasts(PEFs)myogenically into skeletal muscle through overexpression of the pig MYOD1 gene and modulation of the FGF,TGF-β,WNT,and cAMP signaling ***:The MYOD1 overexpression vector was constructed based on comparative sequence analysis,demonstrating that pig MYOD1 has evolutionarily conserved domains across various *** forced MYOD1 expression through these vectors triggered the expression of endogenous muscle markers,transdifferentiated muscle cells from fibroblasts were not ***,various signaling molecules,including FGF2,SB431542,CHIR99021,and forskolin,along with MYOD1 overexpression were applied to enhance the myogenic *** modified conditions led to the derivation of myotubes and activation of muscle markers in PEFs,as determined by qPCR and ***,a sarcomere-like structure was observed,indicating that terminally differentiated skeletal muscle could be obtained from transdifferentiated ***:In summary,we established a protocol for reprogramming MYOD1-overexpressing PEFs into the mature skeletal muscle using signaling *** myogenic reprogramming can be used as a cell source for muscle disease models in regenerative medicine and the production of cultured meat in cellular agriculture.

关键词： MYOD1 Pig Sequence analysis Skeletal muscle transdifferentiation

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Role of Insulin-like Growth Factor II Receptor in transdifferentiation of Free Silica-induced Primary Rat Lung Fibroblasts

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Biomedical and Environmental Sciences 2013年第12期26卷 979-985页

作者： HAO Chang Fu LI Xiao Fang YAO Wu Department of Occupational Health and Occupational Disease College of Public Health Zhengzhou University

Objective To study the role of insulin-like growth factor II receptor in free silica-induced transdifferentiation of primary rat lung fibroblasts Methods Rat lung fibroblasts and rat alveolar macrophages were cultured. A transdifferentiation model of primary rat lung fibroblasts was induced by free silica. Levels of a-SMA protein, IGF-liR protein and mRNA were measured by immunocytochemistry, Western blot and RT-PCR, respectively. Lung fibroblasts were treated with Wortmannin. Results The expression levels of a-SMA concentration and decreased after Wortmann and IGF-IIR increased with the increasing free silica n was used. Conclusion The IGF-IIR plays an important role in free silica-induced transdifferentiation of primary rat lung fibroblasts.

关键词： transdifferentiation Lung fibroblasts Insulin-like growth factor II receptor Silicosis

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In vitro transdifferentiation of corneal epithelial-like cells from human skin-derived precursor cells

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International Journal of Ophthalmology(English edition) 2012年第2期5卷 158-163页

作者： Sarawut Saichanma Ahnond Bunyaratvej Monnipha Sila-asna Department of Pathology Faculty of Medicine Ramathibodi Hospital Mahidol University Bangkok 14140 Thailand Cell Engineering and Tissue Growth Institute of Molecular Biosciences Mahidol University Phuttamonthon Salaya Nakornpathom 73170 Thailand PYY Foundation (Foundation for Sustainable Development) PO Box 19 (kratum ban) Samutakorn 74110 Thailand

The damage of human corneal cells encounter with the problem of availability of corneal cells for replacement. Limitation of the source of corneal cells has been realized. An attempt of development of corneal epithelial-like cells from the human skin-derived precursor (hSKPs) has been made in this study. Combination of three essential growth factors: epidermal growth factor (EGF), keratinocyte growth factor (KGF) and hepatocyte growth factor (HGF) could demonstrate successfully induction of hSKPs to differentiation into corneal cells.The induced cells expressed the appearance of markers of corneal epithelial cells as shown by the presence of keratin 3 (K3) by antibody label and Western blot assay. The K3 gene expression of induced hSKPs cells as shown by reverse transcription-polymerase chain reaction (RT-PCR) technology was also demonstrated. The presence of these markers at both gene and protein levels could lead to our conclusion that the directional transdifferentiation of hSKPs cells into corneal epithelial cells was successfully done under this cell induction protocol. The finding shows a newly available stem cell source can be obtained from easily available skin. Cells from autologous human skin might be used for corneal disorder treatment in future clinical application.

关键词： corneal epithelial-like cell human skin-derived precursor cell transdifferentiation

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Impact of senescence on the transdifferentiation process of human hepatic progenitor-like cells

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World Journal of Stem Cells 2021年第10期13卷 1595-1609页

作者： Francesco Bellanti Giorgia di Bello Rosanna Tamborra Marco Amatruda Aurelio Lo Buglio MichałDobrakowski Aleksandra Kasperczyk Sławomir Kasperczyk Gaetano Serviddio Gianluigi Vendemiale Department of Medical and Surgical Sciences University of FoggiaFoggia 71122Italy Department of Biochemistry Medical University of SilesiaZabrze 41-808Poland

BACKGROUND Senescence is characterized by a decline in hepatocyte function,with impairment of metabolism and regenerative *** models that duplicate liver functions in vitro are essential tools for studying drug metabolism,liver diseases,and organ *** human HepaRG cell line represents an effective model for the study of liver metabolism and hepatic ***,the impact of senescence on HepaRG cells is not yet *** To characterize the effects of senescence on the transdifferentiation capacity and mitochondrial metabolism of human HepaRG *** We compared the transdifferentiation capacity of cells over 10(passage 10[P10])vs *** was evaluated by senescence-associated(SA)beta-galactosidase activity and the comet *** transdifferentiation was analyzed by confocal microscopy and flow cytometry(expression of cluster of differentiation 49a[CD49a],CD49f,CD184,epithelial cell adhesion molecule[EpCAM],and cytokeratin 19[CK19]),quantitative PCR analysis(expression of albumin,cytochrome P4503A4[CYP3A4],γ-glutamyl transpeptidase[γ-GT],and carcinoembryonic antigen[CEA]),and functional analyses(albumin secretion,CYP3A4,andγ-GT).Mitochondrial respiration and the ATP and nicotinamide adenine dinucleotide(NAD^(+))/NAD with hydrogen(NADH)content were also *** SAβ-galactosidase staining was higher in P20 than P10 HepaRG cells;in parallel,the comet assay showed consistent DNA damage in P20 HepaRG *** respect to P10,P20 HepaRG cells exhibited a reduction of CD49a,CD49f,CD184,EpCAM,and CK19 after the induction of ***,lower gene expression of albumin,CYP3A4,andγ-GT,as well as reduced albumin secretion capacity,CYP3A4,andγ-GT activity were reported in transdifferentiated P20 compared to P10 *** contrast,the gene expression level of CEA was not reduced by transdifferentiation in P20 *** note,both cellular and mitochondrial oxygen consumption was lower in P20 than in P10 tra

关键词： Senescence HepaRG cells transdifferentiation Mitochondria Regeneration Nicotinamide adenine dinucleotide

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The Effect of Connective Tissue Growth Factor on Human Renal Tubular Epithelial Cell transdifferentiation

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2004年第4期24卷 350-353页

作者：张春朱忠华邓安国 Department of Nephrology Union Hospital Tongji Medical College Huazhong University of Science and Technology

To investigate the role of connective tissue growth factor (CTGF) in transdifferentiation of human renal tubular epithelial cell (HKC), in vitro cultured HKC cells were divided into 3 groups: negtive control, low dose CTGF-treated group (rh CTGF, 2.5 ng/ml) and high dose CTGF-treated (rhCTGF, 5.0 ng/ml). Then the expression of α-smooth muscle actin (α-SMA) were assessed by indirect immuno-fluorescence, and the percentage of α-SMA positive cells were assessed by flow cytometry. RT-PCR were also performed to examine the mRNA level of α-SMA. Upon the stimulation of different concentrations of rhCTGF, the expression of α-SMA were markedly stronger than that in negative controls. The percentages of α-SMA positive cells were significantly higher in the stimulated groups than that of negative controls (38.9 %, 65.5 % vs 2.4 %, Ptransdifferentiation of human renal tubular epithelial cells towards myofibroblast (Myo-F).

关键词： connective tissue growth factor human renal tubular epithelial cell transdifferentiation

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Effects and mechanisms of total saponins of panax notoginseng on the transdifferentiation of human renal tubular epithelial cells mduced by aristolochic acid Ⅰ

Effects and mechanisms of total saponins of panax notoginsen...

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2008北京国际生理科学学术大会

作者： XU Ya-Ji PAN Qiang-Wen FENG Zhi-Qiang* Department of Physiology Luzhou Medical College Luzhou 646000 China

＜正＞Objective:Aristolochic acid nephropathy（AAN） is a renal disease reported after the introduction of Chinese herbs contained aristolochic acid.A great quantity studies suggest that tubular epithelial-myofibroblast transdifferentiation（TEMT） induced by aristolochic acid（AA） may be the prominent reason of renal interstitial *** saponins of panax notoginseng（PNS） can prevent the development of liver and renal *** objective of our experiment is to observe whether PNS can inhibit AA I triggered TEMT and its mechanism,and to provide a new scientific foundation of blocking chronic aristolochic acid ***:HK-2 cells were divided into 6 groups,including negative control group,PNS control group,AA I induced group and PNS treated group with different doses（200,400,600μg/mL）.Each group was cultured for 48 h in *** morphological change of cultured tubular cells was observed by inverted phase contrast microscopy;the expression ofα-SMA was assessed by immunohistochemistry;the content of TGF-β1 in the culture supernatant was measured by ELISA;the level of CTGF mRNA was evaluated by ***:AA I caused transdifferentiation of HK-2 cells to myofibroblast-like cells and showed strongα-SMA expression,the IOD value ofα-SMA expression was more than that of negative control group,the content of TGF-β1 in supernatant and the level of CTGF mRNA were also increased（P＜0.05）.The morphological change of HK-2 cells was decreased in PNS treated groups with different doses,PNS inhibited the expression ofα-SMA,CTGF mRNA and TGF-β1 in a dose-dependent manner （P＜0.05）.There was no difference in the measured indexes between PNS control group and negative control ***:AA I can increase the content of TGF-β1 and the level of CTGF mRNA,thus increase the expression ofα-SMA and induce TEMT;TEMT andα-SMA expression are down-regulated by PNS in a dosedependent manner,the negative regulation of PNS may be partially achieved by attenuation of TGF-β1

关键词： total saponins of panax notoginseng transdifferentiation α-smooth muscle actin connective tissue growth factor

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Fibroblast transdifferentiation promotes conversion of M1 macrophages and replenishment of cardiac resident macrophages following cardiac injury in mice

Fibroblast transdifferentiation promotes conversion of M1 ma...

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第十四届全国免疫学学术大会

作者： Hongxiang Lv The Affiliated Jiangning Hospital with Nanjing Medical University 江苏大学

Resident cardiac macrophages play important roles in homeostasis, maintenance of cardiac function, and tissue repair. After cardiac injury, monocytes infiltrate the tissue, undergo phenotypic and functional changes, and are involved in inflammatory injury and functional remodelling. However, the fate of cardiac infiltrating/polarized macrophages and the relationship between these cells and resident cardiac macrophage replenishment following injury remain unclear. Our Results showed that angiotensin II(ANG II) induces cardiac fibroblast transdifferentiation into cardiac myofibroblasts(MFBs). In co-cultures with MFBs and murine macrophages, the MFBs promoted macrophage polarization to M1 phenotype, followed by selective apoptosis, which was associated with TNF/TNFR1 axis and independent of NO production. Surprisingly, after 36 h of co-culture, the surviving macrophages were converted to M2 phenotype and settled in heart, which was dependent on leptin produced by MFBs or polarized macrophages via the PI3K or Akt pathway. CCR2CD45.2 cells adoptively transferred into CD45.1 mice with viral myocarditis, differentiated into CD45.2CCR2CX3CR1 M2 cells during the resolution of inflammation and settled within the heart. Our data highlight a novel mechanism related to the renewal or replenishment of cardiac resident macrophages following cardiac injury;and suggest that transdifferentiation of cardiac fibroblasts may promote the resolution of inflammation.

关键词： cardiac resident macrophage transdifferentiation inflammation cardiac injury leptin

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Vasculogenic mimicry of tumor stem/progenitor cells contribute to angiogenesis of gliomas via transdifferentiation

Vasculogenic mimicry of tumor stem/progenitor cells contribu...

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中华医学会神经外科学分会第九次学术会议

作者：董军兰青黄强苏州大学附属第二医院神经外科

Objective Previous studies suggest that tumor cells might be the progenitor for tumor *** vascular tube formation from transdifferentiation of glioma stem/progenitor cells(GSPCs) contribute to angiogenesis of gliomas ... 详细信息

关键词： glioma stem cells transdifferentiation neovascularization

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transdifferentiation of pancreatic α-cells into insulinsecreting cells: From experimental models to underlying mechanisms

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World Journal of Diabetes 2014年第6期5卷 847-853页

作者： Jieli Lu Rami Jaafer Rémy Bonnavion Philippe Bertolino Chang-Xian Zhang Sino-French Life Science and Genomic Center the E-Institute of ShanghaiShanghai Institute of Endocrine and Metabolic DiseasesRuijin HospitalShang-hai Jiao Tong University Shanghai Clinical Center for Endocrine and Metabolic Diseases Ruijin HospitalShanghai Jiao Tong University Inserm U1052 Cancer Research Centre of Lyon CNRS UMR5286 Cancer Research Centre of Lyon Cancer Research Centre of Lyon Université de Lyon

Pancreatic insulin-secreting β-cells are essential regulators of glucose metabolism. New strategies are cur-rently being investigated to create insulin-producing β cells to replace deficient β cells, including the differentiation of either stem or progenitor cells, and the newly uncovered transdifferentiation of mature non-β islet cell types. However, in order to correctly drive any cell to adopt a new β-cell fate, a better understanding of the in vivo mechanisms involved in the plasticity and biology of islet cells is urgently required. Here, we review the recent studies reporting the phenomenon of transdifferentiation of α cells into β cells by focusing on the major candidates and contexts revealed to be involved in adult β-cell regeneration through this process. The possible underlying mechanisms of transdifferentiation and the interactions between several key factors involved in the process are also addressed. We propose that it is of importance to further study the molecular and cellular mechanisms underlying α- to β-cell transdifferentiation, in order to make β-cell regeneration from α cells a relevant and realizable strategy for developing cell-replacement therapy.

关键词： α-cell β-cell transdifferentiation Diabetes mellitus Cell-replacement therapy

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In situ direct reprogramming of astrocytes to neurons via polypyrimidine tract-binding protein 1 knockdown in a mouse model of ischemic stroke

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Neural Regeneration Research 2024年第10期19卷 2240-2248页

作者： Meng Yuan Yao Tang Tianwen Huang Lining Ke En Huang Key Laboratory of Brain Aging and Neurodegenerative Diseases of Fujian Province Fujian Medical UniversityFuzhouFujian ProvinceChina Department of Human Anatomy School of Basic Medical SciencesFujian Medical UniversityFuzhouFujian ProvinceChina Scientific Research Center School of Basic Medical SciencesFujian Medical UniversityFuzhouFujian ProvinceChina Department of Neurology Fujian Medical University Union HospitalFuzhouFujian ProvinceChina Fujian Key Laboratory of Vascular Aging Fujian Medical UniversityFuzhouFujian ProvinceChina

In situ direct reprogramming technology can directly convert endogenous glial cells into functional neurons in vivo for central nervous system repair. Polypyrimidine tract-binding protein 1(PTB) knockdown has been shown to reprogram astrocytes to functional neurons in situ. In this study, we used AAV-PHP.e B-GFAP-sh PTB to knockdown PTB in a mouse model of ischemic stroke induced by endothelin-1, and investigated the effects of GFAP-sh PTB-mediated direct reprogramming to neurons. Our results showed that in the mouse model of ischemic stroke, PTB knockdown effectively reprogrammed GFAP-positive cells to neurons in ischemic foci, restored neural tissue structure, reduced inflammatory response, and improved behavioral function. These findings validate the effectiveness of in situ transdifferentiation of astrocytes, and suggest that the approach may be a promising strategy for stroke treatment.

关键词： astrocyte in situ direct reprogramming ischemic stroke miR-30 based shRNA neuron polypyrimidine tract-binding protein 1 transdifferentiation

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