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RUNX1-205, a novel splice variant of the human RUNX1 gene, has blockage effect on mesoderm-hemogenesis transition and promotion effect during the late stage of hematopoiesis

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Journal of Molecular Cell Biology 2020年第5期12卷 386-396页

作者： Wencui Sun Jiahui Zeng Jing Chang Yuan Xue Yonggang Zhang Xu Pan Ya Zhou Mowen Lai Guohui Bian Qiongxiu Zhou Jiaxing Liu Bo Chen Feng Ma Institute of Blood Transfusion Chinese Academy of Medical Sciences&Peking Union Medical College(CAMS&PUMC)Chengdu 610052China State Key Laboratory of Biotherapy Sichuan UniversityChengdu 61006China State Key Laboratory of Experimental Hematology CAMS&PUMCTianjin 300020China

Runt-related transcription factor 1(RUNX1)is required for definitive hematopoiesis;however,the functions of most human RUNX1 isoforms are *** particular,the effects of RUNX1-205(a novel splice variant that lacks exon 6 in comparison with RUNX1b)on human hematopoiesis are not *** this study,a human embryonic stem cell(hESC)line with inducible RUNX1-205 overexpression was *** of these cells revealed that induction of RUNX1-205 overexpression at early stage did not influence the induction of mesoderm but blocked the emergence of CD34+cells,and the production of hematopoietic stem/progenitor cells was significantly *** addition,the expression of hematopoiesis-related factors was ***,these effects were abolished when RUNX1-205 overexpression was induced after Day 6 in co-cultures of hESCs and AGM-S3 cells,indicating that the inhibitory effect occurred prior to generation of hemogenic endothelial cells,while the promotive effect could be observed during the late stage of *** is very similar to that of ***,the mRNA expression profile of RUNX1-205 during hematopoiesis was distinct from that of RUNX1b,and the protein stability of RUNX1-205 was much higher than that of ***,the function of RUNX1-205 in normal and diseased models should be further explored.

关键词： mesoderm-hemogenesis transition hematopoiesis hematopoietic stem/progenitor cells (HSPCs) RUNX1 mesoderm embryonic stem cells (ESCs)

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Effect of hydroxyurea and etoposide on transduction of human bone marrow mesenchymal stem and progenitor cell by adeno-associated virus vectors

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Acta Pharmacologica Sinica 2004年第2期25卷 70-76页

作者： Xiao-dong JU, Si-quan LOU2, Wei-guo WANG, Jian-qiang PENG3, Hua TIAN Department of Orthopaedics, Peking University Third Hospital, Beijing 100083 3Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100262, China Department of Orthopaedics Peking University Third Hospital Beijing 100083 China

AIM: To study the effect of hydroxyurea and etoposide on transduction of human marrow mesenchymal and progenitor stem cells by adeno-associated virus (AAV). METHODS: Isolated human bone marrow mesenchymal stem and progenitor cells (hMSCs) were cultured in DMEM containing 10 % FBS or 5 % FBS and dexamethasone 1 μmol/L respectively. After being treated with hydroxyurea and etoposide, hMSCs were transduced by AAV-LUC. After two days luciferase activity (relative light unites per second or RLU/s) were tested, which indirectly reflected the relative transduction efficiency of different groups, and virus DNA was isolated by Hirt extraction for Southern hybridization. RESULTS: Transduction luciferase activity and transduction efficiency in cultures treated with hydroxyurea and etoposide were significantly higher than that in control cultures. Dividing cells had about 20-fold higher transduction efficiencycompared with control cells. Transduction efficiency in stationary cells was about 50 times higher than that in control cells. Southern analysis showed that hydroxyurea and etoposide enhanced second-strand DNA synthesis by rAAV. CONCLUSION: Hydroxyurea and etoposide could increase transduction efficiency of hMSCs by AAV vectors, and stationary cells were more sensitive to these drugs than dividing cells.

关键词： simian adenoviruses hydroxyurea etoposide mesoderm stem cells gene transfer techniques

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Tonsil-derived stem cells as a new source of adult stem cells

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World Journal of Stem Cells 2019年第8期11卷 506-518页

作者： Kyung-Ah Cho Hyun Jung Lee Hansaem Jeong Miri Kim Soo Yeon Jung Hae Sang Park Kyung-Ha Ryu Seung Jin Lee Byeongmoon Jeong Hyukjin Lee Han Su Kim Department of Microbiology College of Medicine Ewha Womans University Seoul 07985 South Korea Department of Chemistry and Nano Science Ewha Womans University Seoul 03760 South Korea College of Pharmacy Graduate School of Pharmaceutical Sciences Ewha Womans University Seoul 03760 South Korea Department of Otorhinolaryngology College of Medicine Ewha Womans University Seoul 07985 South Korea Department of Otorhinolaryngology College of Medicine Hallym University Chuncheon 24252 South Korea Department of Pediatrics College of Medicine Ewha Womans University Seoul 07985 South Korea

Located near the oropharynx, the tonsils are the primary mucosal immune organ. Tonsil tissue is a promising alternative source for the high-yield isolation of adult stem cells, and recent studies have reported the identification and isolation of tonsil-derived stem cells (T-SCs) from waste surgical tissue following tonsillectomies in relatively young donors (i.e., under 10 years old). As such, TSCs offer several advantages, including superior proliferation and a shorter doubling time compared to bone marrow-derived mesenchymal stem cells (MSCs). T-SCs also exhibit multi-lineage differentiation, including mesodermal, endodermal (e.g., hepatocytes and parathyroid-like cells), and even ectodermal cells (e.g., Schwann cells). To this end, numbers of researchers have evaluated the practical use of T-SCs as an alternative source of autologous or allogenic MSCs. In this review, we summarize the details of T-SC isolation and identification and provide an overview of their application in cell therapy and regenerative medicine.

关键词： Stem cell Tonsil-derived stem cell Differentiation Endoderm mesoderm Ectoderm Cell therapy

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Overexpression of HOXA9 upregulates NF-κB signaling to promote human hematopoiesis and alter the hematopoietic differentiation potentials

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Cell Regeneration 2021年第1期10卷 95-106页

作者： Jiahui Zeng Danying Yi Wencui Sun Yuanlin Liu Jing Chang Lijiao Zhu Yonggang Zhang Xu Pan Yong Dong Ya Zhou Mowen Lai Guohui Bian Qiongxiu Zhou Jiaxin Liu Bo Chen Feng Ma Chinese Academy of Medical Sciences&Peking Union Medical College(CAMS&PUMC) Institute of Blood TransfusionNo.26Huacai RoadLongtan Industry ParkChenghua DistrictChengdu 610052China State Key Laboratory of Biotherapy Sichuan UniversityChengdu 610065China State Key Laboratory of Experimental Hematology CAMS&PUMCTianjin 300020China

Background: The HOX genes are master regulators of embryogenesis that are also involved inhematopoiesis. HOXA9 belongs to a cluster of HOX genes that play extensively studied roles inhematopoiesis and ***: We established HOXA9-inducible human embryonic stem cells (HOXA9/hESCs) with normalpluripotency and potential for hematopoiesis, which could be used to analyze gene function with highaccuracy. HOXA9/hESCs co-cultured with aorta–gonad–mesonephros-derived stromal cells (AGM-S3) wereinduced to overexpress HOXA9 with doxycycline (DOX) at various times after hematopoiesis started andthen subjected to flow ***: Induction of HOXA9 from Day 4 (D4) or later notably promoted hematopoiesis and also increasedthe production of CD34+ cells and derived populations. The potential for myelogenesis was significantlyelevated while the potential for erythrogenesis was significantly reduced. At D14, a significant promotion ofS phase was observed in green fluorescent protein positive (GFP+) cells overexpressing HOXA9. NF-κBsignaling was also up-regulated at D14 following induction of HOXA9 on D4. All of these effects could becounteracted by addition of an NF-κB inhibitor or siRNA against NFKB1 along with ***: Overexpression of HOXA9 starting at D4 or later during hematopoiesis significantly promotedhematopoiesis and the production of myeloid progenitors while reduced the production of erythroidprogenitors, indicating that HOXA9 plays a key role in hematopoiesis and differentiation of hematopoieticlineages.

关键词： Hematopoiesis Differentiation of hematopoietic lineages HOXA9 mesoderm Human embryonic stem cells(hESCs) Tet-on Inducible expression system

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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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