A Novel Hypothesis and Characterization to Isolate Microvascular Endothelial Cells Simultaneously with AdiposeDerived Stem Cells from the Human Adipose-Derived Stromal Vascular Fraction
10.1007/s13770-021-00332-5
- Author:
Yeon Hee RYU
1
;
Suk-Ho MOON
;
Ki Joo KIM
;
Young Joon JUN
;
Deuk Young OH
;
Sang-Heon KIM
;
Jong-Won RHIE
Author Information
1. Department of Biomedicine and Health Sciences, College of Medicine, The Catholic University of Korea, 222 Banpodaero, Seocho-gu, Seoul 06591, Republic of Korea
- Publication Type:O RI G I N A L A R T I C L E
- From:
Tissue Engineering and Regenerative Medicine
2021;18(3):429-440
- CountryRepublic of Korea
- Language:English
-
Abstract:
BACKGROUND:Angiogenesis and vasculogenesis are essential processes for successful tissue regeneration in tissue engineering and regenerative medicine. The adipose-derived stromal vascular fraction (SVF) is not only a source of adipose stem cells (ASC) but also a suitable source of microvascular endothelial cells because it is a rich capillary network. So, we propose a new hypothesis for isolating adipose-derived human microvascular endothelial cells (HMVEC-A) from the SVF and developed a dual isolation system that isolates two cell types from one tissue.METHOD: To isolate HMVEC-A, we analyzed the supernatant discarded when ASC is isolated from the adipose-derived SVF. Based on this analysis, we assumed that the SVF adherent to the bottom of the culture plate was divided into two fractions: the stromal fraction as the ASC-rich fraction, and the vascular fraction (VF) as the endothelial cells-rich fraction floating in the culture supernatant. VF isolation was optimized and the efficiency was compared, and the endothelial cells characteristics of HMVEC-A were confirmed by flow cytometric analysis, immunocytochemistry (ICC), a DiI-acetylated low-density lipoprotein (DiI-Ac-LDL) uptake, and in vitro tube formation assay.
RESULTS:Consistent with the hypothesis, we found a large population of HMVEC-A in the VF and isolated these HMVEC-A by our isolation method. Additionally, this method had higher yields and shorter doubling times than other endothelial cells isolation methods and showed typical morphological and phenotypic characteristics of endothelial cells.
CONCLUSION:Cells obtained by the method according to our hypothesis can be applied as a useful source for studies such as tissue-to-tissue networks, angiogenesis and tissue regeneration, patient-specific cell therapy, and organoid chips.
