Expression of ID2 in lung bud organoids promotes pulmonary vascular formation
Objective To investigate the role of inhibitor of DNA binding-2 (ID2) in pulmonary vasculature formation in early lung development. Methods CRISPR/Cas9 system was used to edit ID2 in human embryonic stem cells (hESCs) to construct ID2 knockout strains, hESCs and ID2 knockout strains were further d...
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| Format: | Article |
| Language: | zho |
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Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.
2021-05-01
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| Series: | Jichu yixue yu linchuang |
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| Online Access: | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/a210013.pdf |
| _version_ | 1827390587665383424 |
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| author | LIU Hong-xian, YANG Jun |
| author_facet | LIU Hong-xian, YANG Jun |
| author_sort | LIU Hong-xian, YANG Jun |
| collection | DOAJ |
| description | Objective To investigate the role of inhibitor of DNA binding-2 (ID2) in pulmonary vasculature formation in early lung development. Methods CRISPR/Cas9 system was used to edit ID2 in human embryonic stem cells (hESCs) to construct ID2 knockout strains, hESCs and ID2 knockout strains were further differentiated into endothelial cells(ECs) or lung bud organoids (LBOs).Quantitative PCR was used to detect ID2, CD31, NKX2.1 and other genes expression during differentiation, flow cytometry was performed to analyze the differentiation efficiency, Western blot was applied to check ID2, CD31 and PAX9 protein expression, 3D co-culture of ECs and LBOs was performed to compare the effect of LBOs derived from different hESCs on the tubular formation of ECs. Results ID2 promoted cells to enter the mesoderm stage during the differentiation of hESCs into ECs, but ID2 deletion led to the formation of a large number of ECs with tubular defects. ID2 promoted the differentiation of hESCs into LBOs.In the 3D co-culture system, LBOs could promote ECs tube-forming ability, while the deficiency of ID2 weakened this promotion. Conclusions In early stage of pulmonary tissue development, ID2 affects the differenti- ation and function of ECs and LBOs, loss of ID2 in lung bud reduces the angiogenesis of surrounded ECs. |
| first_indexed | 2024-03-08T16:53:53Z |
| format | Article |
| id | doaj.art-5d3ce9eec30e43b18bae0f548f9cb0bf |
| institution | Directory Open Access Journal |
| issn | 1001-6325 |
| language | zho |
| last_indexed | 2024-03-08T16:53:53Z |
| publishDate | 2021-05-01 |
| publisher | Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College. |
| record_format | Article |
| series | Jichu yixue yu linchuang |
| spelling | doaj.art-5d3ce9eec30e43b18bae0f548f9cb0bf2024-01-05T03:05:08ZzhoInstitute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.Jichu yixue yu linchuang1001-63252021-05-01415623629Expression of ID2 in lung bud organoids promotes pulmonary vascular formationLIU Hong-xian, YANG Jun01. State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005; ;2. Department of Physiology, School of Basic Medical Sciences,Zhejiang University School of Medicine, Hangzhou 310058, ChinaObjective To investigate the role of inhibitor of DNA binding-2 (ID2) in pulmonary vasculature formation in early lung development. Methods CRISPR/Cas9 system was used to edit ID2 in human embryonic stem cells (hESCs) to construct ID2 knockout strains, hESCs and ID2 knockout strains were further differentiated into endothelial cells(ECs) or lung bud organoids (LBOs).Quantitative PCR was used to detect ID2, CD31, NKX2.1 and other genes expression during differentiation, flow cytometry was performed to analyze the differentiation efficiency, Western blot was applied to check ID2, CD31 and PAX9 protein expression, 3D co-culture of ECs and LBOs was performed to compare the effect of LBOs derived from different hESCs on the tubular formation of ECs. Results ID2 promoted cells to enter the mesoderm stage during the differentiation of hESCs into ECs, but ID2 deletion led to the formation of a large number of ECs with tubular defects. ID2 promoted the differentiation of hESCs into LBOs.In the 3D co-culture system, LBOs could promote ECs tube-forming ability, while the deficiency of ID2 weakened this promotion. Conclusions In early stage of pulmonary tissue development, ID2 affects the differenti- ation and function of ECs and LBOs, loss of ID2 in lung bud reduces the angiogenesis of surrounded ECs.http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/a210013.pdfinhibitor of dna binding-2 (id2)|lung bud organoids(lbos)|endothelial cells(ecs) |
| spellingShingle | LIU Hong-xian, YANG Jun Expression of ID2 in lung bud organoids promotes pulmonary vascular formation Jichu yixue yu linchuang inhibitor of dna binding-2 (id2)|lung bud organoids(lbos)|endothelial cells(ecs) |
| title | Expression of ID2 in lung bud organoids promotes pulmonary vascular formation |
| title_full | Expression of ID2 in lung bud organoids promotes pulmonary vascular formation |
| title_fullStr | Expression of ID2 in lung bud organoids promotes pulmonary vascular formation |
| title_full_unstemmed | Expression of ID2 in lung bud organoids promotes pulmonary vascular formation |
| title_short | Expression of ID2 in lung bud organoids promotes pulmonary vascular formation |
| title_sort | expression of id2 in lung bud organoids promotes pulmonary vascular formation |
| topic | inhibitor of dna binding-2 (id2)|lung bud organoids(lbos)|endothelial cells(ecs) |
| url | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/a210013.pdf |
| work_keys_str_mv | AT liuhongxianyangjun expressionofid2inlungbudorganoidspromotespulmonaryvascularformation |