Enhanced HSC-like cell generation from mouse pluripotent stem cells in a 3D induction system cocultured with stromal cells
Abstract Background Decades of efforts have attempted to differentiate the pluripotent stem cells (PSCs) into truly functional hematopoietic stem cells (HSCs), yet the problems of low differentiation efficiency in vitro and poor hematopoiesis reconstitution in vivo still exist, mainly attributing to...
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| Language: | English |
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BMC
2021-06-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-021-02434-2 |
| _version_ | 1819138567826309120 |
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| author | Wei Shan Qin Yu Yan Long Qian Luo Honghu Li Yingli Han Yulin Xu Shan Fu Xiangjun Zeng Cong Wei Yang Gao Xiaoqing Li Xia Li Lifei Zhang Lizhen Liu Ming Chen Pengxu Qian He Huang |
| author_facet | Wei Shan Qin Yu Yan Long Qian Luo Honghu Li Yingli Han Yulin Xu Shan Fu Xiangjun Zeng Cong Wei Yang Gao Xiaoqing Li Xia Li Lifei Zhang Lizhen Liu Ming Chen Pengxu Qian He Huang |
| author_sort | Wei Shan |
| collection | DOAJ |
| description | Abstract Background Decades of efforts have attempted to differentiate the pluripotent stem cells (PSCs) into truly functional hematopoietic stem cells (HSCs), yet the problems of low differentiation efficiency in vitro and poor hematopoiesis reconstitution in vivo still exist, mainly attributing to the lack of solid, reproduced, or pursued differentiation system. Methods In this study, we established an in vitro differentiation system yielding in vivo hematopoietic reconstitution hematopoietic cells from mouse PSCs through a 3D induction system followed by coculture with OP9 stromal cells. The in vivo hematopoietic reconstitution potential of c-kit+ cells derived from the mouse PSCs was evaluated via m-NSG transplantation assay. Flow cytometry analysis, RNA-seq, and cell cycle analysis were used to detect the in vitro hematopoietic ability of endothelial protein C receptor (EPCR, CD201) cells generated in our induction system. Results The c-kit+ cells from 3D self-assembling peptide induction system followed by the OP9 coculture system possessed apparently superiority in terms of in vivo repopulating activity than that of 3D induction system followed by the 0.1% gelatin culture. We interestingly found that our 3D+OP9 system enriched a higher percentage of CD201+c-kit+cells that showed more similar HSC-like features such as transcriptome level and CFU formation ability than CD201-c-kit+cells, which have not been reported in the field of mouse PSCs hematopoietic differentiation. Moreover, CD201+ hematopoietic cells remained in a relatively slow cycling state, consistent with high expression levels of P57 and Ccng2. Further, we innovatively demonstrated that notch signaling pathway is responsible for in vitro CD201+ hematopoietic cell induction from mouse PSCs. Conclusions Altogether, our findings lay a foundation for improving the efficiency of hematopoietic differentiation and generating in vivo functional HSC-like cells from mouse PSCs for clinical application. |
| first_indexed | 2024-12-22T11:08:50Z |
| format | Article |
| id | doaj.art-75c3bbcec1854169b9ac8cdf5a801612 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-22T11:08:50Z |
| publishDate | 2021-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-75c3bbcec1854169b9ac8cdf5a8016122022-12-21T18:28:13ZengBMCStem Cell Research & Therapy1757-65122021-06-0112111710.1186/s13287-021-02434-2Enhanced HSC-like cell generation from mouse pluripotent stem cells in a 3D induction system cocultured with stromal cellsWei Shan0Qin Yu1Yan Long2Qian Luo3Honghu Li4Yingli Han5Yulin Xu6Shan Fu7Xiangjun Zeng8Cong Wei9Yang Gao10Xiaoqing Li11Xia Li12Lifei Zhang13Lizhen Liu14Ming Chen15Pengxu Qian16He Huang17Bone Marrow Transplantation Center, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityCollege of Life Science, Zhejiang Chinese Medical UniversityBone Marrow Transplantation Center, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityBone Marrow Transplantation Center, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityBone Marrow Transplantation Center, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityBone Marrow Transplantation Center, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityBone Marrow Transplantation Center, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityBone Marrow Transplantation Center, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityBone Marrow Transplantation Center, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityBone Marrow Transplantation Center, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityDepartment of Hematology, Sir Run Run Shaw Hospital, Zhejiang University School of MedicineBone Marrow Transplantation Center, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityBone Marrow Transplantation Center, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityDepartment of Hematology, Sir Run Run Shaw Hospital, Zhejiang University School of MedicineBone Marrow Transplantation Center, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityInstitute of Hematology, Zhejiang UniversityInstitute of Hematology, Zhejiang UniversityBone Marrow Transplantation Center, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityAbstract Background Decades of efforts have attempted to differentiate the pluripotent stem cells (PSCs) into truly functional hematopoietic stem cells (HSCs), yet the problems of low differentiation efficiency in vitro and poor hematopoiesis reconstitution in vivo still exist, mainly attributing to the lack of solid, reproduced, or pursued differentiation system. Methods In this study, we established an in vitro differentiation system yielding in vivo hematopoietic reconstitution hematopoietic cells from mouse PSCs through a 3D induction system followed by coculture with OP9 stromal cells. The in vivo hematopoietic reconstitution potential of c-kit+ cells derived from the mouse PSCs was evaluated via m-NSG transplantation assay. Flow cytometry analysis, RNA-seq, and cell cycle analysis were used to detect the in vitro hematopoietic ability of endothelial protein C receptor (EPCR, CD201) cells generated in our induction system. Results The c-kit+ cells from 3D self-assembling peptide induction system followed by the OP9 coculture system possessed apparently superiority in terms of in vivo repopulating activity than that of 3D induction system followed by the 0.1% gelatin culture. We interestingly found that our 3D+OP9 system enriched a higher percentage of CD201+c-kit+cells that showed more similar HSC-like features such as transcriptome level and CFU formation ability than CD201-c-kit+cells, which have not been reported in the field of mouse PSCs hematopoietic differentiation. Moreover, CD201+ hematopoietic cells remained in a relatively slow cycling state, consistent with high expression levels of P57 and Ccng2. Further, we innovatively demonstrated that notch signaling pathway is responsible for in vitro CD201+ hematopoietic cell induction from mouse PSCs. Conclusions Altogether, our findings lay a foundation for improving the efficiency of hematopoietic differentiation and generating in vivo functional HSC-like cells from mouse PSCs for clinical application.https://doi.org/10.1186/s13287-021-02434-23D systemPluripotent stem cellsHematopoiesisCD201Notch |
| spellingShingle | Wei Shan Qin Yu Yan Long Qian Luo Honghu Li Yingli Han Yulin Xu Shan Fu Xiangjun Zeng Cong Wei Yang Gao Xiaoqing Li Xia Li Lifei Zhang Lizhen Liu Ming Chen Pengxu Qian He Huang Enhanced HSC-like cell generation from mouse pluripotent stem cells in a 3D induction system cocultured with stromal cells Stem Cell Research & Therapy 3D system Pluripotent stem cells Hematopoiesis CD201 Notch |
| title | Enhanced HSC-like cell generation from mouse pluripotent stem cells in a 3D induction system cocultured with stromal cells |
| title_full | Enhanced HSC-like cell generation from mouse pluripotent stem cells in a 3D induction system cocultured with stromal cells |
| title_fullStr | Enhanced HSC-like cell generation from mouse pluripotent stem cells in a 3D induction system cocultured with stromal cells |
| title_full_unstemmed | Enhanced HSC-like cell generation from mouse pluripotent stem cells in a 3D induction system cocultured with stromal cells |
| title_short | Enhanced HSC-like cell generation from mouse pluripotent stem cells in a 3D induction system cocultured with stromal cells |
| title_sort | enhanced hsc like cell generation from mouse pluripotent stem cells in a 3d induction system cocultured with stromal cells |
| topic | 3D system Pluripotent stem cells Hematopoiesis CD201 Notch |
| url | https://doi.org/10.1186/s13287-021-02434-2 |
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