Chemically defined generation of human definitive hematopoietic stem and progenitor cells
Summary: Here, we present a protocol to efficiently direct human pluripotent stem cells (hPSCs) into hematopoietic stem and progenitor cells (HSPCs) under a chemically defined, albumin-free system. We describe the induction of aorta-gonad-mesonephros-like hematopoiesis from hPSCs into SOX17+ hemogen...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-03-01
|
| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722008334 |
| _version_ | 1798005238918021120 |
|---|---|
| author | Juhyung Jung Yun Chang Gyuhyung Jin Qing Deng Xiaojun Lance Lian Xiaoping Bao |
| author_facet | Juhyung Jung Yun Chang Gyuhyung Jin Qing Deng Xiaojun Lance Lian Xiaoping Bao |
| author_sort | Juhyung Jung |
| collection | DOAJ |
| description | Summary: Here, we present a protocol to efficiently direct human pluripotent stem cells (hPSCs) into hematopoietic stem and progenitor cells (HSPCs) under a chemically defined, albumin-free system. We describe the induction of aorta-gonad-mesonephros-like hematopoiesis from hPSCs into SOX17+ hemogenic endothelium and then into CD34+CD45+ HSPCs via application of Wnt activator and TGFβ inhibitor, respectively. The generated HSPCs, characterized by flow cytometry and colony-forming unit assay, express definitive hematopoiesis markers and exhibit multilineage differentiation potential and the capacity to expand.For complete details on the use and execution of this protocol, please refer to Chang et al. (2022a, 2022b).1,2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-11T12:36:01Z |
| format | Article |
| id | doaj.art-cddbe11e3994499b849abb81df110f02 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-11T12:36:01Z |
| publishDate | 2023-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-cddbe11e3994499b849abb81df110f022022-12-22T04:23:37ZengElsevierSTAR Protocols2666-16672023-03-0141101953Chemically defined generation of human definitive hematopoietic stem and progenitor cellsJuhyung Jung0Yun Chang1Gyuhyung Jin2Qing Deng3Xiaojun Lance Lian4Xiaoping Bao5Davdidson School of Chemical Engineering, Purdue University, West Lafayette, IN 47906, USA; Purdue University Center for Cancer Research, West Lafayette, IN 47906, USADavdidson School of Chemical Engineering, Purdue University, West Lafayette, IN 47906, USA; Purdue University Center for Cancer Research, West Lafayette, IN 47906, USA; Corresponding authorDavdidson School of Chemical Engineering, Purdue University, West Lafayette, IN 47906, USA; Purdue University Center for Cancer Research, West Lafayette, IN 47906, USAPurdue University Center for Cancer Research, West Lafayette, IN 47906, USA; Department of Biological Sciences, Purdue University, West Lafayette, IN 47907, USADepartment of Biomedical Engineering, The Huck Institutes of the Life Sciences, Department of Biology, Pennsylvania State University, University Park, PA 16802, USADavdidson School of Chemical Engineering, Purdue University, West Lafayette, IN 47906, USA; Purdue University Center for Cancer Research, West Lafayette, IN 47906, USA; Corresponding authorSummary: Here, we present a protocol to efficiently direct human pluripotent stem cells (hPSCs) into hematopoietic stem and progenitor cells (HSPCs) under a chemically defined, albumin-free system. We describe the induction of aorta-gonad-mesonephros-like hematopoiesis from hPSCs into SOX17+ hemogenic endothelium and then into CD34+CD45+ HSPCs via application of Wnt activator and TGFβ inhibitor, respectively. The generated HSPCs, characterized by flow cytometry and colony-forming unit assay, express definitive hematopoiesis markers and exhibit multilineage differentiation potential and the capacity to expand.For complete details on the use and execution of this protocol, please refer to Chang et al. (2022a, 2022b).1,2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166722008334Cell BiologyCell cultureCell-based AssaysStem CellsCell DifferentiationBiotechnology and bioengineering |
| spellingShingle | Juhyung Jung Yun Chang Gyuhyung Jin Qing Deng Xiaojun Lance Lian Xiaoping Bao Chemically defined generation of human definitive hematopoietic stem and progenitor cells STAR Protocols Cell Biology Cell culture Cell-based Assays Stem Cells Cell Differentiation Biotechnology and bioengineering |
| title | Chemically defined generation of human definitive hematopoietic stem and progenitor cells |
| title_full | Chemically defined generation of human definitive hematopoietic stem and progenitor cells |
| title_fullStr | Chemically defined generation of human definitive hematopoietic stem and progenitor cells |
| title_full_unstemmed | Chemically defined generation of human definitive hematopoietic stem and progenitor cells |
| title_short | Chemically defined generation of human definitive hematopoietic stem and progenitor cells |
| title_sort | chemically defined generation of human definitive hematopoietic stem and progenitor cells |
| topic | Cell Biology Cell culture Cell-based Assays Stem Cells Cell Differentiation Biotechnology and bioengineering |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722008334 |
| work_keys_str_mv | AT juhyungjung chemicallydefinedgenerationofhumandefinitivehematopoieticstemandprogenitorcells AT yunchang chemicallydefinedgenerationofhumandefinitivehematopoieticstemandprogenitorcells AT gyuhyungjin chemicallydefinedgenerationofhumandefinitivehematopoieticstemandprogenitorcells AT qingdeng chemicallydefinedgenerationofhumandefinitivehematopoieticstemandprogenitorcells AT xiaojunlancelian chemicallydefinedgenerationofhumandefinitivehematopoieticstemandprogenitorcells AT xiaopingbao chemicallydefinedgenerationofhumandefinitivehematopoieticstemandprogenitorcells |