Rapid isolation of adipose tissue-derived stem cells by the storage of lipoaspirates

Yonsei Med J. 2011 Nov;52(6):999-1007. doi: 10.3349/ymj.2011.52.6.999.

Abstract

Purpose: This study examined a rapid isolation method decreasing the time and cost of the clinical application of adipose tissue-derived stem cells (ASCs).

Materials and methods: Aliquots (10 g) of the lipoaspirates were stored at 4°C without supplying oxygen or nutrients. At the indicated time points, the yield of mononuclear cells was evaluated and the stem cell population was counted by colony forming unit-fibroblast assays. Cell surface markers, stem cell-related transcription factors, and differentiation potentials of ASCs were analyzed.

Results: When the lipoaspirates were stored at 4°C, the total yield of mononuclear cells decreased, but the stem cell population was enriched. These ASCs expressed CD44, CD73, CD90, CD105, and HLA-ABC but not CD14, CD31, CD34, CD45, CD117, CD133, and HLA-DR. The number of ASCs increased 1×10(14) fold for 120 days. ASCs differentiated into osteoblasts, adipocytes, muscle cells, or neuronal cells.

Conclusion: ASCs isolated from lipoaspirates and stored for 24 hours at 4°C have similar properties to ASCs isolated from fresh lipoaspirates. Our results suggest that ASCs can be isolated with high frequency by optimal storage at 4°C for 24 hours, and those ASCs are highly proliferative and multipotent, similar to ASCs isolated from fresh lipoaspirates. These ASCs can be useful for clinical application because they are time- and cost-efficient, and these cells maintain their stemness for a long time, like ASCs isolated from fresh lipoaspirates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5'-Nucleotidase / metabolism
  • Adipose Tissue / cytology*
  • Adult
  • Antigens, CD / metabolism
  • Cell Differentiation / physiology
  • Cells, Cultured
  • Endoglin
  • Female
  • Humans
  • Hyaluronan Receptors / metabolism
  • Immunoblotting
  • Immunohistochemistry
  • Immunophenotyping
  • Mesenchymal Stem Cells / metabolism
  • Muscle Development / genetics
  • Muscle Development / physiology
  • Osteogenesis / genetics
  • Osteogenesis / physiology
  • Receptors, Cell Surface / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stem Cells / cytology*
  • Stem Cells / metabolism
  • Thy-1 Antigens / metabolism
  • Young Adult

Substances

  • Antigens, CD
  • ENG protein, human
  • Endoglin
  • Hyaluronan Receptors
  • Receptors, Cell Surface
  • Thy-1 Antigens
  • 5'-Nucleotidase