Efficient endothelial and smooth muscle cell differentiation from human pluripotent stem cells through a simplified insulin-free culture system

Zhang, Fengzhi; Zhu, Yonglin; Chen, Jing; Kuang, Wenhua; Huang, Rujin; Duan, Fuyu; Li, Yaqian; Wang, Lin; Qiu, Hui; Chen, Xia; Jia, Ming; Liu, Peng; Du, Yanan; Chang, Sophia Chia-Ning; Chen, Ligong; Na, Jie

doi:10.1016/j.biomaterials.2021.120713

Cited by 14 publications

(16 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Interestingly, insulin was reported to suppress the arterial marker gene expression 55 . Insulin withdrawal significantly increases arterial ECs formation at the cost of HPC 9 , 19 . Therefore, optimizing insulin concentration and cell cycle adjustment may help produce arterial HECs with lymphoid differentiation and definitive hematopoiesis potential.…”

Section: Discussionmentioning

confidence: 98%

See 1 more Smart Citation

Integrative epigenomic and transcriptomic analysis reveals the requirement of JUNB for hematopoietic fate induction

et al. 2022

Self Cite

View full text Add to dashboard Cite

Human pluripotent stem cell differentiation towards hematopoietic progenitor cell can serve as an in vitro model for human embryonic hematopoiesis, but the dynamic change of epigenome and transcriptome remains elusive. Here, we systematically profile the chromatin accessibility, H3K4me3 and H3K27me3 modifications, and the transcriptome of intermediate progenitors during hematopoietic progenitor cell differentiation in vitro. The integrative analyses reveal sequential opening-up of regions for the binding of hematopoietic transcription factors and stepwise epigenetic reprogramming of bivalent genes. Single-cell analysis of cells undergoing the endothelial-to-hematopoietic transition and comparison with in vivo hemogenic endothelial cells reveal important features of in vitro and in vivo hematopoiesis. We find that JUNB is an essential regulator for hemogenic endothelium specialization and endothelial-to-hematopoietic transition. These studies depict an epigenomic roadmap from human pluripotent stem cells to hematopoietic progenitor cells, which may pave the way to generate hematopoietic progenitor cells with improved developmental potentials.

show abstract

Section: Discussionmentioning

confidence: 98%

“…In the in vitro differentiation system, HPC development occurs through sequential stages. HPSCs are induced to mesoderm cells with strong vascular differentiation potential 9 . Endothelial progenitor cells are then induced from these mesoderm cells under the stimulus of VEGF and bFGF 10 .…”

Section: Introductionmentioning

confidence: 99%

Integrative epigenomic and transcriptomic analysis reveals the requirement of JUNB for hematopoietic fate induction

et al. 2022

Self Cite

View full text Add to dashboard Cite

show abstract

“…Inspired by human embryonic development, several research groups have developed protocols to differentiate hPSCs into HPs through the derivation of an in vitro hemogenic endothelium [5,[18][19][20][21]. Other groups have reported efficient differentiation of hPSCs into functional ECs [22][23][24][25][26][27][28][29][30][31][32][33]. In both cases, protocols require the selection of endothelial progenitors at around day 6 of differentiation.…”

Section: Introductionmentioning

confidence: 99%

Endothelial and hematopoietic hPSCs differentiation via a hematoendothelial progenitor

et al. 2022

View full text Add to dashboard Cite

Background hPSC-derived endothelial and hematopoietic cells (ECs and HCs) are an interesting source of cells for tissue engineering. Despite their close spatial and temporal embryonic development, current hPSC differentiation protocols are specialized in only one of these lineages. In this study, we generated a hematoendothelial population that could be further differentiated in vitro to both lineages. Methods Two hESCs and one hiPSC lines were differentiated into a hematoendothelial population, hPSC-ECs and blast colonies (hPSC-BCs) via CD144+-embryoid bodies (hPSC-EBs). hPSC-ECs were characterized by endothelial colony-forming assay, LDL uptake assay, endothelial activation by TNF-α, nitric oxide detection and Matrigel-based tube formation. Hematopoietic colony-forming cell assay was performed from hPSC-BCs. Interestingly, we identified a hPSC-BC population characterized by the expression of both CD144 and CD45. hPSC-ECs and hPSC-BCs were analyzed by flow cytometry and RT-qPCR; in vivo experiments have been realized by ischemic tissue injury model on a mouse dorsal skinfold chamber and hematopoietic reconstitution in irradiated immunosuppressed mouse from hPSC-ECs and hPSC-EB-CD144+, respectively. Transcriptomic analyses were performed to confirm the endothelial and hematopoietic identity of hESC-derived cell populations by comparing them against undifferentiated hESC, among each other’s (e.g. hPSC-ECs vs. hPSC-EB-CD144+) and against human embryonic liver (EL) endothelial, hematoendothelial and hematopoietic cell subpopulations. Results A hematoendothelial population was obtained after 84 h of hPSC-EBs formation under serum-free conditions and isolated based on CD144 expression. Intrafemorally injection of hPSC-EB-CD144+ contributed to the generation of CD45+ human cells in immunodeficient mice suggesting the existence of hemogenic ECs within hPSC-EB-CD144+. Endothelial differentiation of hPSC-EB-CD144+ yields a population of > 95% functional ECs in vitro. hPSC-ECs derived through this protocol participated at the formation of new vessels in vivo in a mouse ischemia model. In vitro, hematopoietic differentiation of hPSC-EB-CD144+ generated an intermediate population of > 90% CD43+ hPSC-BCs capable to generate myeloid and erythroid colonies. Finally, the transcriptomic analyses confirmed the hematoendothelial, endothelial and hematopoietic identity of hPSC-EB-CD144+, hPSC-ECs and hPSC-BCs, respectively, and the similarities between hPSC-BC-CD144+CD45+, a subpopulation of hPSC-BCs, and human EL hematopoietic stem cells/hematopoietic progenitors. Conclusion The present work reports a hPSC differentiation protocol into functional hematopoietic and endothelial cells through a hematoendothelial population. Both lineages were proven to display characteristics of physiological human cells, and therefore, they represent an interesting rapid source of cells for future cell therapy and tissue engineering.

show abstract

“…There, ECs keep playing an important role in controlling proliferation and differentiation of HSCs (17). Despite these close developmental and functional relationship between ECs and HCs, most of the current differentiation protocols of hPSCs focus on the derivation on one of these lineages (5,(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35). Other authors differentiated both lineages (ECs and HCs) (36,37).…”

Section: Introductionmentioning

confidence: 99%

Endothelial and hematopoietic hPSCs differentiation via an hematoendothelial progenitor.

Vargas-Valderrama

Ponsen

Oberlin

et al. 2022

Preprint

View full text Add to dashboard Cite

Background: hPSC-derived endothelial and hematopoietic cells (ECs and HCs) are an interesting source of cells for tissue engineering. Despite their close spatial and temporal embryonic development, current hPSC differentiation protocols are specialized in only one of these lineages. In this study, we generated a hematoendothelial population that could be further differentiated in vitro to both lineages. Methods: Two hESCs and one hiPSC lines were differentiated into a hematoendothelial population, hPSC-ECs and blast colonies (hPSC-BCs) via CD144+-embryoid bodies (hPSC-EBs). hPSC-ECs were characterized by endothelial colony forming assay, LDL uptake assay, endothelial activation by TNF-α, Nitric Oxyde detection and Matrigel-based tube formation. Hematopoietic colony forming cell assay was performed from hPSC-BCs. Interestingly, we identified a hPSC-BC population characterized by the expression of both CD144 and CD45. hPSC-ECs and hPSC-BCs were analyzed by flow cytometry and RT-qPCR, in vivo experiments have been realized by ischemic tissue injury model on a mouse dorsal skinfold chamber and hematopoietic reconstitution in irradiated immunosuppressed mouse from hPSC-ECs and hPSC-EB-CD144+ respectively. Transcriptomic analyses were performed to confirm the endothelial and hematopoietic identity of hESC-derived cell populations by comparing them against undifferentiated hESC, among each other’s (e.g., hPSC-ECs vs hPSC-EB-CD144+) and against human embryonic liver (EL) endothelial, hematoendothelial and hematopoietic cell sub populations.Results: A hematoendothelial population was obtained after 84 hours of hPSC-EBs formation under serum-free conditions and isolated based on CD144 expression. Intrafemorally injection of hPSC-EB-CD144+ contributed to the generation of CD45+ human cells in immunodeficient mice suggesting the existence of hemogenic ECs within hPSC-EB-CD144+. Endothelial differentiation of hPSC-EB-CD144+ yields a population of >95% functional ECs in vitro. hPSC-ECs derived through this protocol participated at the formation of new vessels in vivo in a mouse ischemia model. In vitro, hematopoietic differentiation of hPSC-EB-CD144+ generated an intermediate population of >90% CD43+ hPSC-BCs capable to generate myeloid and erythroid colonies. Finally, the transcriptomic analyses confirmed the hematoendothelial, endothelial and hematopoietic identity of hPSC-EB-CD144+, hPSC-ECs and hPSC-BCs, respectively, and the similarities between hPSC-BC-CD144+CD45+, a subpopulation of hPSC-BCs, and human EL hematopoietic stem cells/hematopoietic progenitors. Conclusion: The present work reports a hPSC differentiation protocol into functional hematopoietic and endothelial cells through a hematoendothelial population. Both lineages were proven to display characteristics of physiological human cells and therefore, they represent an interesting rapid source of cells for future cell therapy and tissue engineering.

show abstract

Efficient endothelial and smooth muscle cell differentiation from human pluripotent stem cells through a simplified insulin-free culture system

Cited by 14 publications

References 49 publications

Integrative epigenomic and transcriptomic analysis reveals the requirement of JUNB for hematopoietic fate induction

Integrative epigenomic and transcriptomic analysis reveals the requirement of JUNB for hematopoietic fate induction

Endothelial and hematopoietic hPSCs differentiation via a hematoendothelial progenitor

Endothelial and hematopoietic hPSCs differentiation via an hematoendothelial progenitor.

Contact Info

Product

Resources

About