title: Development of fully defined xeno-free culture system for the preparation and propagation of cell therapy-compliant human adipose stem cells.
abstract: BACKGROUND
  Adipose tissue is an attractive and abundant source of multipotent stem cells. Human adipose stem cells (ASCs) have shown to have therapeutic relevancy in diverse clinical applications. Nevertheless, expansion of ASCs is often necessary before performing clinical studies. Standard in vitro cell-culture techniques use animal-derived reagents that should be avoided in clinical use because of safety issues. Therefore, xeno- and serum-free (XF/SF) reagents are highly desirable for enhancing the safety and quality of the transplanted ASCs.
  
  
  METHODS
  In the current study, animal component-free isolation and cell-expansion protocols were developed for ASCs. StemPro MSC SFM XF medium with either CELLstart™ CTS™ coating or Coating Matrix Kit were tested for their ability to support XF/SF growth. Basic stem-cell characteristics such as immunophenotype (CD3, CD11a, CD14, CD19, CD34, CD45RO, CD54, CD73, CD80, CD86, CD90, CD105, HLA-DR), proliferation, and differentiation potential were assessed in XF/SF conditions and compared with human serum (HS) or traditionally used fetal bovine serum (FBS) cultures.
  
  
  RESULTS
  ASCs cultured in XF/SF conditions had significantly higher proliferation rates compared with HS/FBS cultures. Characteristic immunophenotypes of ASCs were maintained in every condition; however, cells expanded in XF/SF conditions showed significantly lower expression of CD54 (intercellular adhesion molecule 1, ICAM-1) at low passage number. Further, multilineage differentiation potential of ASCs was maintained in every culture condition.
  
  
  CONCLUSIONS
  Our findings demonstrated that the novel XF/SF conditions maintained the basic stem cell features of ASCs and the animal-free workflow followed in this study has great potential in clinical cell therapies.
authors.forename: Mimmi
authors.initials: M
authors.lastname: Patrikoski
authors.forename: Miia
authors.initials: M
authors.lastname: Juntunen
authors.forename: Shayne
authors.initials: S
authors.lastname: Boucher
authors.forename: Andrew
authors.initials: A
authors.lastname: Campbell
authors.forename: Mohan C
authors.initials: MC
authors.lastname: Vemuri
authors.forename: Bettina
authors.initials: B
authors.lastname: Mannerström
authors.forename: Susanna
authors.initials: S
authors.lastname: Miettinen
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000328:Adult; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D064987:Cell- and Tissue-Based Therapy; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000187:drug effects; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D005260:Female; D006801:Humans; D016130:Immunophenotyping; D008875:Middle Aged; D010012:Osteogenesis / Q000187:drug effects; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D016895:Culture Media, Serum-Free