Clonal isolation of endothelial colony-forming cells from early gestation chorionic villi of human placenta for fetal tissue regeneration

Abstract

BACKGROUND Endothelial colony-forming cells (ECFCs) have been implicated in the process of vascularization, which includes vasculogenesis and angiogenesis. Vasculogenesis is a de novo formation of blood vessels, and is an essential physiological process that occurs during embryonic development and tissue regeneration. Angiogenesis is the growth of new capillaries from pre-existing blood vessels, which is observed both prenatally and postnatally. The placenta is an organ composed of a variety of fetal-derived cells, including ECFCs, and therefore has significant potential as a source of fetal ECFCs for tissue engineering. AIM To investigate the possibility of isolating clonal ECFCs from human early gestation chorionic villi (CV-ECFCs) of the placenta, and assess their potential for tissue engineering. METHODS The early gestation chorionic villus tissue was dissociated by enzyme digestion. Cells expressing CD31 were selected using magnetic-activated cell sorting, and plated in endothelial-specific growth medium. After 2-3 wks in culture, colonies displaying cobblestone-like morphology were manually picked using cloning cylinders. We characterized CV-ECFCs by flow cytometry, immunophenotyping, Data sharing statement: We agree to share the research methods and data in this study to public.

Bioactive extracellular matrix scaffolds engineered with proangiogenic proteoglycan mimetics and loaded with endothelial progenitor cells promote neovascularization and diabetic wound healing

“…Additionally, MSCs have also demonstrated the ability to regulate the immune system through multiple independent pathways [ 38 ] which may facilitate wound healing. On the other hand, EPCs can engraft and become part of the neovasculature if the environment is permissive [ [39] , [40] , [41] ]. EPCs could also secrete angiogenic growth factors that may facilitate wound healing [ 42 ].…”

Biological Behavioral Alterations of the Post-neural Differentiated Dental Pulp Stem Cells Through an in situ Microenvironment

“…However, CD105 and CD146 expression was lower than that observed in the control group; CD105 and CD146 expression was decreased from 99.37 to 58.41% and from 97.60 to 69.75%, respectively. CD146 and CD105 are both primarily endothelial cell markers associated with vascular endothelial cell activity and angiogenesis ( Nakashima et al, 2009 ; Gao et al, 2020 ). These results again suggested that d-DPSCs cultured in 10% FBS α-MEM had the ability to restore partly their stemness but had less capacity for angiogenesis.…”

Developing an Injectable Nanofibrous Extracellular Matrix Hydrogel With an Integrin αvβ3 Ligand to Improve Endothelial Cell Survival, Engraftment and Vascularization

“…All lentiviral constructs were generated at the UC Davis Institute for Regenerative Cures (IRC) Vector Core. To track the cell fate and behavior in vitro and in vivo , ECFCs were transduced with the pCCLc-MNDU3-LUC-PGK-EGFP-WPRE vector as previously described ( Kumar et al, 2018 ; Gao et al, 2019 , 2020 ; Rose et al, 2020 ). Transduction was performed in transduction medium consisting of DMEM high glucose (HyClone), 10% FBS (HyClone), and 8 μg/mL protamine sulfate (MP Biomedicals) for 6 h. The vectors were transduced at a multiplicity of infection (MOI) of 10.…”