Differentiating Human Pluripotent Stem Cells to Vascular Endothelial Cells for Regenerative Medicine, Tissue Engineering, and Disease Modeling

Bertucci, Taylor; Kakarla, Shravani; Kim, Diana; Dai, Guohao

doi:10.1007/978-1-0716-1708-3_1

Cited by 6 publications

(5 citation statements)

References 20 publications

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“…The data contradicted these previously published results in the sense that TIE2 had been intensely expressed at the endothelial level both in small mesenchymal vessels and in the aortic endothelium at the level of the thoracic aorta in the seven-week embryo. Other studies on TIE2 expression at the endothelial level have been conducted on an animal model [ 28 ], mainly avian, but not on human embryonic tissue. In the avian model, TIE2 was expressed at the level of the endothelium of embryonic veins and very weakly and inconsistently at the level of arterial endothelium.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Vasculogenic Factors in the Developing Embryo at Weeks Five and Seven With a Special Focus on CD133 and TIE2 Markers

Tomescu,

Sas,

Sarb

et al. 2024

Cureus

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BackgroundHuman embryo vasculogenesis (blood vessel development starting from endothelial precursors) includes the ability of mesenchymal cells and pluripotent stem cells to differentiate into endothelial cells. Quantification of endothelial progenitor cells is difficult to assess during the early steps of human embryo development due to several factors, especially due to the paucity of human embryo tissue which is usually discarded after early-stage pregnancy abortive methods. CD133 (Promimin-1) is a general marker of progenitor cells, but combined with other endothelial markers such as CD34, it may identify endothelial progenitor cells during embryonic development. CD34 immunohistochemistry was previously performed by our team to identify human embryo capillaries and comparatively assess microvessel density between different human embryonic tissues. TIE2 is an angiopoietin receptor strongly involved in the newly formed blood vessel maturation due to its expression in some mesenchymal precursors for future pericytes. CD34 assesses the presence of endothelial cells but its single use does not evaluate the endothelial progenitor state as CD133 may do nor vessel maturation as TIE2 may do. Data about the dynamics of CD133/TIE2 expression in the early stages of human embryo development are scarce. Hence, in this study, we aimed to comparatively assess the dynamic of CD133+ endothelial precursors and TIE2 expression on five and seven-week-old human embryonic tissues with a special emphasis on their expression on embryonic vascular beds. MethodologyCD133 and TIE2 immunohistochemistry was performed on five and seven-week-old human embryonic tissues followed by their quantification using the Qu Path digital image analysis (DIA) automated method. ResultsCD133 and TIE2 showed divergent patterns of expression during the initial phases of human embryonic development, specifically in the vascular endothelium of tiny capillaries. The expression of CD133 in endothelial cells lining the perfused lumen gradually decreased from five to seven-week-old embryos. It remained expressed with greater intensity in cells located at the tip of the vascular bud that emerged into pre-existing capillaries. TIE2 was much more specific than CD133, being restricted to the level of the vascular endothelium; therefore, it was easier to quantify using digital image analysis. The endothelium of the embryonic aorta was an exception to the divergent expression, as CD133 and TIE2 were consistently coexpressed in the seven-week-old embryo. The Qu Path DIA assessment increased the accuracy of CD133 and TIE2 evaluation, being the first time they were quantified by using automated software and not manually. ConclusionsHigh heterogeneity of CD133 and TIE2 was observed between five and seven-week-old embryonic tissues as well as between different embryonic regions from the same gestational age. The unique finding of CD133/TIE2 co-expression persistence inside aortic endothelium needs further studies to elucidate the role of this co-expression.

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Section: Discussionmentioning

confidence: 99%

Evaluation of Vasculogenic Factors in the Developing Embryo at Weeks Five and Seven With a Special Focus on CD133 and TIE2 Markers

Tomescu,

Sas,

Sarb

et al. 2024

Cureus

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“…Once a stable iPSC line is created, EC differentiation is typically driven using small molecules and growth factors. 32 , 33 In transdifferentiation, EC reprogramming is driven using a mix of overexpressed transcription factors and small molecules. 2 , 21 , 22 , 34 In summary, full iPSC reprogramming is time consuming, costly, and inefficient, while transdifferentiation can occur much more rapidly.…”

Section: Discussionmentioning

confidence: 99%

“… 38 , 39 iPSC-EC differentiation is met with greatly varying levels of efficiency (6%–16%, 35 18%, 37 >60% 31 ), with higher efficiencies coming out in more recent years. 32 , 33 These iPSC-ECs can express key EC proteins VE-CAD and CD31, as well as vWF. They also take up and retain LDL, respond to inflammatory stimuli, and form tubules when cultured on Matrigel.…”

Section: Discussionmentioning

confidence: 99%

SOX17/ETV2 improves the direct reprogramming of adult fibroblasts to endothelial cells

Grath,

Dai

2024

Cell Reports Methods

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“…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

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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.

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Differentiating Human Pluripotent Stem Cells to Vascular Endothelial Cells for Regenerative Medicine, Tissue Engineering, and Disease Modeling

Cited by 6 publications

References 20 publications

Evaluation of Vasculogenic Factors in the Developing Embryo at Weeks Five and Seven With a Special Focus on CD133 and TIE2 Markers

Evaluation of Vasculogenic Factors in the Developing Embryo at Weeks Five and Seven With a Special Focus on CD133 and TIE2 Markers

SOX17/ETV2 improves the direct reprogramming of adult fibroblasts to endothelial cells

Endothelial and hematopoietic hPSCs differentiation via a hematoendothelial progenitor

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