Clonal isolation of endothelial colony-forming cells from early gestation chorionic villi of human placenta for fetal tissue regeneration
Open accessAbstract
BACKGROUND: 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, tube formation assay, and Dil-Ac-LDL uptake assay. Viral transduction of CV-ECFCs was performed using a Luciferase/tdTomato-containing lentiviral vector, and transduction efficiency was tested by fluorescent microscopy and flow cytometry. Compatibility of CV-ECFCs with a delivery vehicle was determined using an FDA approved, small intestinal submucosa extracellular matrix scaffold. RESULTS: CV-ECFCs using 100 mg of early gestational chorionic villus tissue. Immunophenotypic analyses by flow cytometry demonstrated that CV-ECFCs highly expressed the endothelial markers CD31, CD144, CD146, CD105, CD309, only partially expressed CD34, and did not express CD45 and CD90. CV-ECFCs were capable of acetylated low-density lipoprotein uptake and tube formation, similar to cord blood-derived ECFCs (CB-ECFCs). CV-ECFCs can be transduced with a Luciferase/tdTomato-containing lentiviral vector at a transduction efficiency of 85.1%. Seeding CV-ECFCs on a small intestinal submucosa extracellular matrix scaffold confirmed that CV-ECFCs were compatible with the biomaterial scaffold. CONCLUSION: In summary, we established a magnetic sorting-assisted clonal isolation approach to derive CV-ECFCs. A substantial number of CV-ECFCs can be obtained within a short time frame, representing a promising novel source of ECFCs for fetal treatments.
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ECFCs sourced from cord blood or placenta can create de novo vascular networks in vitro and maintain the capacity to integrate with host vasculature in vivo.
“Endothelial colony forming cells (ECFC), sourced from cord‐blood or the placenta, demonstrate the ability to create de novo vascular networks in vitro, whilst maintaining the capacity to integrate with host vasculature in vivo.[ 46 , 47 , 48 ]”
From Coordinated Development of Immune Cell Populations in Vascularized Skin Organoids from Human Induced Pluripotent Stem Cells (2025) · cites this paper as [47]
The Results section ('CV-ECFCs are functionally comparable to CB-ECFCs'; page unavailable) reports that placenta- and cord-blood-derived ECFCs formed similar tubular structures in vitro. The Introduction says ECFCs can integrate into the circulatory system, but this paper does not demonstrate that its placental ECFCs integrate with host vasculature in vivo. That part of the claim is therefore not established for both cell sources. (LLM, conf=0.84)
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MethodSelf-citation
- Authors
- Gao, K., He, S., Kumar, P., Farmer, D., Zhou, J., & Wang, A.
- Affiliations
- Shriners Hospitals for Children - Northern California (US)
- Corresponding author
- Wang, Aijun
- Venue
- World Journal of Stem Cells
- Publisher
- Baishideng Publishing Group Inc.
- Published
- 2020-02-26
- Volume
- 12
- Issue
- 2
- Pages
- 123-138
- ISSN
- 1948-0210 (ISSN-L)
- PMC
- PMC7062038
- OpenAlex
- W3008433274
- Cited by
- 17
- References
- 51
- Citations by year
- 2026: 1 · 2025: 3 · 2024: 2 · 2023: 2 · 2022: 2 · 2021: 2 · 2020: 5
- FWCI
- 0.89
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- cc-by-nc
- Locations
- World Journal of Stem Cells (publishedVersion, cc-by-nc)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- eScholarship (California Digital Library) (submittedVersion)
- Funders
- March of Dimes Foundation (85108-NCA-19, 5R01NS100761-02, R03HD091601-01, 5FY1682, 87200-NCA-19)
- California Institute of Regenerative Medicine (11404, CLIN1-11404, 85108-NCA-19, PC1-08103)
- Shriners Hospitals for Children (85119-NCA-18, 5R01NS100761, 87200-NCA-19, R03HD091601-01, 5FY1682, 84704‐NCA‐19, 5R01NS100761-02, 85120‐NCA‐16, 85108‐NCA‐19, CLIN1-11404, 87200)
- National Institutes of Health (CLIN1-11404, 85108-NCA-19, 87200-NCA-19)
- University of California, Davis
- National Institute of Neurological Disorders and Stroke (R01NS100761)
- Eunice Kennedy Shriver National Institute of Child Health and Human Development (R03HD091601)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Angiogenesis and VEGF in Cancer
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
- Keywords
- endothelial colony-forming cells, chorionic villi, human placenta, clonal isolation, angiogenesis, endothelial markers, lentiviral transduction, extracellular matrix scaffolds, tissue engineering, magnetic cell separation, tube formation, acetylated low-density lipoprotein uptake, viral transduction
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