High-efficient serum-free differentiation of endothelial cells from human iPS cells
Abstract Introduction Endothelial cells (ECs) form the inner lining of all blood vessels of the body play important roles in vascular tone regulation, hormone secretion, anticoagulation, regulation of blood cell adhesion and immune cell extravasation. Limitless ECs sources are required to further in...
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| Format: | Article |
| Language: | English |
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BMC
2022-06-01
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| Series: | Stem Cell Research & Therapy |
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| Online Access: | https://doi.org/10.1186/s13287-022-02924-x |
| _version_ | 1818469588740866048 |
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| author | Sarkawt Hamad Daniel Derichsweiler John Antonydas Gaspar Konrad Brockmeier Jürgen Hescheler Agapios Sachinidis Kurt Paul Pfannkuche |
| author_facet | Sarkawt Hamad Daniel Derichsweiler John Antonydas Gaspar Konrad Brockmeier Jürgen Hescheler Agapios Sachinidis Kurt Paul Pfannkuche |
| author_sort | Sarkawt Hamad |
| collection | DOAJ |
| description | Abstract Introduction Endothelial cells (ECs) form the inner lining of all blood vessels of the body play important roles in vascular tone regulation, hormone secretion, anticoagulation, regulation of blood cell adhesion and immune cell extravasation. Limitless ECs sources are required to further in vitro investigations of ECs’ physiology and pathophysiology as well as for tissue engineering approaches. Ideally, the differentiation protocol avoids animal-derived components such as fetal serum and yields ECs at efficiencies that make further sorting obsolete for most applications. Method Human induced pluripotent stem cells (hiPSCs) are cultured under serum-free conditions and induced into mesodermal progenitor cells via stimulation of Wnt signaling for 24 h. Mesodermal progenitor cells are further differentiated into ECs by utilizing a combination of human vascular endothelial growth factor A165 (VEGF), basic fibroblast growth factor (bFGF), 8-Bromoadenosine 3′,5′-cyclic monophosphate sodium salt monohydrate (8Bro) and melatonin (Mel) for 48 h. Result This combination generates hiPSC derived ECs (hiPSC-ECs) at a fraction of 90.9 ± 1.5% and is easily transferable from the two-dimensional (2D) monolayer into three-dimensional (3D) scalable bioreactor suspension cultures. hiPSC-ECs are positive for CD31, VE-Cadherin, von Willebrand factor and CD34. Furthermore, the majority of hiPSC-ECs express the vascular endothelial marker CD184 (CXCR4). Conclusion The differentiation method presented here generates hiPSC-ECs in only 6 days, without addition of animal sera and at high efficiency, hence providing a scalable source of hiPSC-ECs. |
| first_indexed | 2024-04-13T21:26:39Z |
| format | Article |
| id | doaj.art-1e160f6c5ed2454b8d053c9df50c2f23 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-04-13T21:26:39Z |
| publishDate | 2022-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-1e160f6c5ed2454b8d053c9df50c2f232022-12-22T02:29:18ZengBMCStem Cell Research & Therapy1757-65122022-06-0113111610.1186/s13287-022-02924-xHigh-efficient serum-free differentiation of endothelial cells from human iPS cellsSarkawt Hamad0Daniel Derichsweiler1John Antonydas Gaspar2Konrad Brockmeier3Jürgen Hescheler4Agapios Sachinidis5Kurt Paul Pfannkuche6Medical Faculty, Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of CologneMedical Faculty, Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of CologneMedical Faculty, Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of CologneDepartment of Pediatric Cardiology, University Hospital of CologneMedical Faculty, Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of CologneMedical Faculty, Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of CologneMedical Faculty, Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of CologneAbstract Introduction Endothelial cells (ECs) form the inner lining of all blood vessels of the body play important roles in vascular tone regulation, hormone secretion, anticoagulation, regulation of blood cell adhesion and immune cell extravasation. Limitless ECs sources are required to further in vitro investigations of ECs’ physiology and pathophysiology as well as for tissue engineering approaches. Ideally, the differentiation protocol avoids animal-derived components such as fetal serum and yields ECs at efficiencies that make further sorting obsolete for most applications. Method Human induced pluripotent stem cells (hiPSCs) are cultured under serum-free conditions and induced into mesodermal progenitor cells via stimulation of Wnt signaling for 24 h. Mesodermal progenitor cells are further differentiated into ECs by utilizing a combination of human vascular endothelial growth factor A165 (VEGF), basic fibroblast growth factor (bFGF), 8-Bromoadenosine 3′,5′-cyclic monophosphate sodium salt monohydrate (8Bro) and melatonin (Mel) for 48 h. Result This combination generates hiPSC derived ECs (hiPSC-ECs) at a fraction of 90.9 ± 1.5% and is easily transferable from the two-dimensional (2D) monolayer into three-dimensional (3D) scalable bioreactor suspension cultures. hiPSC-ECs are positive for CD31, VE-Cadherin, von Willebrand factor and CD34. Furthermore, the majority of hiPSC-ECs express the vascular endothelial marker CD184 (CXCR4). Conclusion The differentiation method presented here generates hiPSC-ECs in only 6 days, without addition of animal sera and at high efficiency, hence providing a scalable source of hiPSC-ECs.https://doi.org/10.1186/s13287-022-02924-xHuman induced pluripotent stem cellsiPS cellshiPSCsDifferentiationEndothelial cellsRegenerative medicine, 2D monolayer culture |
| spellingShingle | Sarkawt Hamad Daniel Derichsweiler John Antonydas Gaspar Konrad Brockmeier Jürgen Hescheler Agapios Sachinidis Kurt Paul Pfannkuche High-efficient serum-free differentiation of endothelial cells from human iPS cells Stem Cell Research & Therapy Human induced pluripotent stem cells iPS cells hiPSCs Differentiation Endothelial cells Regenerative medicine, 2D monolayer culture |
| title | High-efficient serum-free differentiation of endothelial cells from human iPS cells |
| title_full | High-efficient serum-free differentiation of endothelial cells from human iPS cells |
| title_fullStr | High-efficient serum-free differentiation of endothelial cells from human iPS cells |
| title_full_unstemmed | High-efficient serum-free differentiation of endothelial cells from human iPS cells |
| title_short | High-efficient serum-free differentiation of endothelial cells from human iPS cells |
| title_sort | high efficient serum free differentiation of endothelial cells from human ips cells |
| topic | Human induced pluripotent stem cells iPS cells hiPSCs Differentiation Endothelial cells Regenerative medicine, 2D monolayer culture |
| url | https://doi.org/10.1186/s13287-022-02924-x |
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