title: Fast isolation and expansion of multipotent cells from adipose tissue based on chitosan-selected primary culture.
abstract: Adipose-derived adult stem cells (ASCs) have gained much attention because of their multipotency and easy access. Here we describe a novel chitosan-based selection (CS) system instead of the conventional plastic adherence (PA) to obtain the primary ASCs. The minimal amount of adipose tissue for consistent isolation of ASCs is reduced from 10 mL to 5 mL. The selection is based on the specific interaction between cells and chitosan materials, which separate ASCs by forming spheroids during primary culture. The primary culture period was reduced from 4 days to one day and more ASCs (ten-fold expansion) were achieved in a week. The average duration for obtaining 1 × 10(7) cells takes about seven days from 5 mL of adipose tissue, compared to 14 days using the conventional PA method from 10 mL of adipose tissue. The replicative senescence of CS-ASCs is not evident until the fifteenth passage (vs. eighth for the PA-ASCs). The obtained ASCs (CS-ASCs) have less doubling time for the same passage of cells and show greater stemness than those obtained from the conventional PA method (PA-ASCs). Moreover, CS-ASCs undergo trilineage differentiation more effectively than PA-ASCs. The greater differentiation potential of CS-ASCs may be associated with the enrichment and maintenance of CD271 positive cells by chitosan selection of primary culture.
authors.affiliation: Institute of Polymer Science and Engineering, National Taiwan University, Taipei, Taiwan.
authors.forename: Guo-Shiang
authors.initials: GS
authors.lastname: Huang
authors.affiliation: Institute of Polymer Science and Engineering, National Taiwan University, Taipei, Taiwan.
authors.forename: Ting-Chen
authors.initials: TC
authors.lastname: Tseng
authors.affiliation: Division of Plastic and Reconstructive Surgery, Department of Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.
authors.forename: Niann-Tzyy
authors.initials: NT
authors.lastname: Dai
authors.affiliation: Division of Plastic and Reconstructive Surgery, Department of Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.
authors.forename: Keng-Yen
authors.initials: KY
authors.lastname: Fu
authors.affiliation: Department of Orthopedics, Shuang Ho Hospital, Taipei Medical University, Taipei, Taiwan.
authors.forename: Lien-Guo
authors.initials: LG
authors.lastname: Dai
authors.affiliation: Institute of Polymer Science and Engineering, National Taiwan University, Taipei, Taiwan; Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, Taipei, Taiwan. Electronic address: shhsu@ntu.edu.tw.
authors.forename: Shan-Hui
authors.initials: SH
authors.lastname: Hsu
mesh_terms: D000068816:Adapalene / Q000032:analysis; D000273:Adipose Tissue / Q000166:cytology*; D053687:Adult Stem Cells / Q000166:cytology*; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D018929:Cell Culture Techniques / Q000191:economics / Q000379:methods*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000191:economics / Q000379:methods*; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry*; D039902:Multipotent Stem Cells / Q000166:cytology*; D011817:Rabbits; D013997:Time Factors
chemical_list: D001672:Biocompatible Materials; D000068816:Adapalene; D048271:Chitosan
keywords: Adipose-derived adult stem cells (ASCs); CD271; Chitosan; Plastic adherence (PA)