title: Isolation of adipose-derived stem cells by using a subfractionation culturing method.
abstract: OBJECTIVE
  Adipose-derived stem cells (ASCs) isolated from subcutaneous adipose tissue have been tested in clinical trials. However, ASCs isolated by enzyme digestion and centrifugation are heterogeneous and exhibit wide variation in regenerative potential and clinical outcomes. Therefore, we developed a new method for isolating clonal ASCs (cASCs) that does not use enzyme digestion or centrifugation steps.
  
  
  METHODS
  In addition to cell surface markers and differentiation potential, we compared the mitogenic, paracrine and hair growth-promoting effects of ASCs isolated by the gradient centrifugation method (GCM) or by the new subfractionation culturing method (SCM).
  
  
  RESULTS
  We selected three cASCs isolated by SCM that showed high rates of proliferation. The cell surface markers expressed by ASCs isolated by GCM or SCM were very similar, and SCM-isolated ASCs could potentially differentiate into different cell lineages. However, cASC lines exhibited better mitogenic and paracrine effects than ASCs isolated by GCM. The expression of Diras3, Myb, Cdca7, Mki67, Rrm2, Cdk1 and Ccna2, which may play a key role in cASC proliferation, was upregulated in cASCs. In addition, cASCs exhibited enhanced hair growth-promoting effects in dermal papilla cells and animal experiments.
  
  
  CONCLUSIONS
  SCM generates a highly homogeneous population of ASCs via a simple and effective procedure that can be used in therapeutic settings.
authors: Inha University School of Medicine, Translational Research Center and Inha Research Institute for Medical Sciences , 7-206, 3-Ga, Shinheung-Dong, Chung-Gu, Inchon 400-711 , Korea +82 32 890 2460 ; +82 32 890 2462 ; sunuksong@inha.ac.kr.
authors: TacGhee
authors: T
authors: Yi
authors: Wang-Kyun
authors: WK
authors: Kim
authors: Joon-Seok
authors: JS
authors: Choi
authors: Seung Yong
authors: SY
authors: Song
authors: Juhee
authors: J
authors: Han
authors: Ji Hye
authors: JH
authors: Kim
authors: Won-Serk
authors: WS
authors: Kim
authors: Sang Gyu
authors: SG
authors: Park
authors: Hyun-Joo
authors: HJ
authors: Lee
authors: Yun Kyoung
authors: YK
authors: Cho
authors: Sung-Joo
authors: SJ
authors: Hwang
authors: Sun U
authors: SU
authors: Song
authors: Jong-Hyuk
authors: JH
authors: Sung
mesh_terms: D017667:Adipocytes / Q000502:physiology*; D000273:Adipose Tissue / Q000166:cytology* / Q000502:physiology*; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D019070:Cell Lineage; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D051379:Mice; D008809:Mice, Inbred C3H; D013234:Stem Cells / Q000166:cytology
keywords: adipose-derived stem cell; hair regeneration; isolation; proliferation; subfractionation culturing method