title: Human endometrial stromal stem cells differentiate into megakaryocytes with the ability to produce functional platelets.
abstract: Human endometrium is a high dynamic tissue that contains endometrial stromal stem cells (hESSCs). The hESSCs have been differentiated into a number of cell lineages. However, differentiation of hESSCs into megakaryocytes (MKs) has not yet been investigated. The aim of this study was to investigate the feasibility of MK generation from hESSCs and subsequent production of functional platelets (PLTs). In our study, hESSCs were cultured from endometrial stromal cells as confirmed by positive stromal cell specific markers (CD90 and CD29) and negative hematopoietic stem cell markers (CD45 and CD34) expression. Then, hESSCs were differentiated in a medium supplemented with thrombopoietin (TPO) for 18 days. The MK differentiation was analyzed by flow cytometry and confocal microscopy. The differentiation medium was collected for PLT production analysis by flow cytometry, transmission electron microscopy and functional measurements. Our results show: 1) MKs were successfully generated from hESSCs as identified by expression of specific markers (CD41a: 1 ± 0.09% and 39 ± 3.0%; CD42b: 1.2 ± 0.06% and 28 ± 2.0%, control vs. differentiation) accompanied with reduction of pluripotent transcription factors (Oct4 and Sox2) expression; 2) The level of PLTs in the differentiation medium was 16 ± 1 number/µl as determined by size (2-4 µm) and CD41a expression (CD41a: 1 ± 0.4% and 90±2.0%, control vs. differentiation); 3) Generated PLTs were functional as evidenced by the up-regulation of CD62p expression and fibrinogen binding following thrombin stimulation; 4) Released PLTs showed similar ultra-structure characteristics (alpha granules, vacuoles and dense tubular system) as PLTs from peripheral blood determined by electron microscopic analysis. Data demonstrate the feasibility of generating MKs from hESSCs, and that the generated MKs release functional PLTs. Therefore, hESSCs could be a potential new stem cell source for in vitro MK/PLT production.
authors.affiliation: Department of Pharmacology & Toxicology, Boonshoft School of Medicine, Wright State University, Dayton, Ohio, USA.
authors.forename: Jinju
authors.initials: J
authors.lastname: Wang
authors.forename: Shuzhen
authors.initials: S
authors.lastname: Chen
authors.forename: Cheng
authors.initials: C
authors.lastname: Zhang
authors.forename: Samantha
authors.initials: S
authors.lastname: Stegeman
authors.forename: Teresa
authors.initials: T
authors.lastname: Pfaff-Amesse
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Wenfeng
authors.initials: W
authors.lastname: Zhang
authors.forename: Lawrence
authors.initials: L
authors.lastname: Amesse
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Chen
mesh_terms: D000328:Adult; D000818:Animals; D001792:Blood Platelets / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D002454:Cell Differentiation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D004717:Endometrium / Q000166:cytology*; D005260:Female; D005340:Fibrinogen / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D008533:Megakaryocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008875:Middle Aged; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D019007:P-Selectin / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects; D013917:Thrombin / Q000494:pharmacology
chemical_list: D003470:Culture Media; D050814:Octamer Transcription Factor-3; D019007:P-Selectin; C494173:POU5F1 protein, human; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D005340:Fibrinogen; D013917:Thrombin