Non-enzymatic isolation followed by supplementation of basic fibroblast growth factor improves proliferation, clonogenic capacity and SSEA-4 expression of perivascular cells from human umbilical cord

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• 作者：Borim An (1)
  Sunghun Na (2)
  Sungeun Lee (3)
  Woo Jin Kim (1)
  Se-Ran Yang (4) (6)
  Heung-Myong Woo (5) (6)
  Songyi Kook (2)
  Yoonki Hong (1)
  Haengseok Song (3)
  Seok-Ho Hong (1) (6)
  
  1. Department of Internal Medicine ; School of Medicine ; Kangwon National University ; 1 Kangwondaehak-gil ; Chuncheon-si ; Gangwon-do ; 200-701 ; Republic of Korea
  2. Department of Obstetrics and Gynecology ; School of Medicine ; Kangwon National University ; Chuncheon ; Republic of Korea
  3. Department of Biomedical Science ; CHA University ; Seoul ; Republic of Korea
  4. Department of Thoracic and Cardiovascular Surgery ; School of Medicine ; Kangwon National University ; Chuncheon ; Republic of Korea
  6. Stem Cell Institute ; Kangwon National University ; Chuncheon ; Republic of Korea
  5. College of Veterinary Medicine ; Kangwon National University ; Chuncheon ; Republic of Korea
  
• 关键词：Perivascular cells ; bFGF ; Non ; enzymatic isolation ; Umbilical cord ; SSEA ; 4
• 刊名：Cell and Tissue Research
• 出版年：2015
• 出版时间：March 2015
• 年：2015
• 卷：359
• 期：3
• 页码：767-777
• 全文大小：2,944 KB
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• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Human Genetics
  Proteomics
  Molecular Medicine
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0878

文摘

Multipotent perivascular cells (PVCs) have recently gained attention as an alternative source for cell-based regenerative medicine. Because of their rarity in human tissues, the development of efficient methods to isolate and expand PVCs from various fetal and adult tissues is necessary to obtain a clinically relevant number of cells that maintain progenitor potency. We report a simple non-enzymatic isolation (NE) method of PVCs from human umbilical cord (HUC) and compare its efficiency with the conventional collagenase treatment method (CT) in terms of proliferation, immunophenotype, clonogenic capacity, and differentiation potential. Cells isolated by NE expressed the accepted surface marker profile of PVCs and possessed multilineage differentiation potential. Whereas both methods provided similar patterns or levels of immunophenotypes and proliferation, PVCs obtained by NE maintained a higher level of CD146(+) frequency compared with that of CT over passages and displayed greater in vitro osteogenic differentiation potential and clonogenic capacity than CT-PVCs. We assess the potential of various exogenous factors to boost the proliferation of NE- and CT-PVCs in vitro. Supplementation of basic fibroblast growth factor (bFGF) provided optimal conditions that significantly enhanced their proliferation rate. This treatment drove the cells into S phase and increased the proportion of stage-specific antigen-4-positive population without altering other immunophenotypes. Thus, the NE method with bFGF supplementation offers an alternative way for obtaining sufficient numbers of HUCPVCs that have good clonogenic and differentiation potential and that are applicable at therapeutic doses for regenerative medicine.