The Effect of Storage Time on Adipose-Derived Stem Cell Recovery from Human Lipoaspirates

Multipotent adipose-derived stromal/stem cells (ASCs) can be isolated with high yield from human subcutaneous lipoaspirates. This study reports our experience isolating, expanding, differentiating and immunophenotypically characterizing ASCs over a period of 4 days after having surgically obtained the lipoaspirate samples. The ultimate goal is to understand how to optimize the consistent isolation of ASCs from lipoaspirates. The length of time between adipose tissue harvest and processing will need to be systematically evaluated with respect to cell yield, viability, and function since some distance is likely to exist between the plastic surgeon’s office where lipoaspiration is performed and the current Good Manufacturing Practices (cGMP) laboratory where the ASCs are isolated. The objective of this study was to determine the effect of time delays on the yield and function of ASCs after collagenase digestion. We were able to isolate ASCs from lipoaspirates up to 72 h after the surgical procedure. The ASCs isolated on sequential days after the original tissue harvest proliferated, differentiated and maintained cell surface markers. We found that the initial 24-hour period is optimal for isolating ASCs with respect to cell yield and that there was no significant difference between ASC cell proliferation and differentiation ability within this period of time. In contrast, each of these parameters declined significantly for tissues maintained at room temperature for 48 or 72 h prior to isolation. These findings should be considered in the future development of standard operating procedures for cGMP processing of clinical-grade human ASCs.

[1]  D. A. Gomes,et al.  Endothelial Differentiation of Human Stem Cells Seeded onto Electrospun Polyhydroxybutyrate/Polyhydroxybutyrate-Co-Hydroxyvalerate Fiber Mesh , 2012, PloS one.

[2]  R. Reis,et al.  Distinct Stem Cells Subpopulations Isolated from Human Adipose Tissue Exhibit Different Chondrogenic and Osteogenic Differentiation Potential , 2011, Stem Cell Reviews and Reports.

[3]  J. Gimble,et al.  Yield and characterization of subcutaneous human adipose-derived stem cells by flow cytometric and adipogenic mRNA analyzes. , 2010, Cytotherapy.

[4]  D. Machin,et al.  Phase II Clinical Trials , 2010 .

[5]  K. Harii,et al.  Progenitor‐Enriched Adipose Tissue Transplantation as Rescue for Breast Implant Complications , 2010, The breast journal.

[6]  R. Suuronen,et al.  Novel maxillary reconstruction with ectopic bone formation by GMP adipose stem cells. , 2009, International journal of oral and maxillofacial surgery.

[7]  R. Reis,et al.  Adipose tissue-derived stem cells and their application in bone and cartilage tissue engineering. , 2009, Tissue engineering. Part B, Reviews.

[8]  D. García-Olmo,et al.  Expanded Adipose-Derived Stem Cells for the Treatment of Complex Perianal Fistula: a Phase II Clinical Trial , 2009, Diseases of the colon and rectum.

[9]  K. Harii,et al.  Cell‐Assisted Lipotransfer for Facial Lipoatrophy: Efficacy of Clinical Use of Adipose‐Derived Stem Cells , 2008, Dermatologic surgery : official publication for American Society for Dermatologic Surgery [et al.].

[10]  E. Ravussin,et al.  Isolation of human adipose-derived stem cells from biopsies and liposuction specimens. , 2008, Methods in molecular biology.

[11]  D. García-Olmo,et al.  Expanded adipose-derived stem cells for the treatment of complex perianal fistula including Crohn's disease. , 2008, Expert opinion on biological therapy.

[12]  D. García-Olmo,et al.  Treatment of enterocutaneous fistula in Crohn’s Disease with adipose-derived stem cells: a comparison of protocols with and without cell expansion , 2008, International Journal of Colorectal Disease.

[13]  K. Yoshimura,et al.  Influences of Preservation at Various Temperatures on Liposuction Aspirates , 2007, Plastic and reconstructive surgery.

[14]  K. Harii,et al.  Cell-Assisted Lipotransfer for Cosmetic Breast Augmentation: Supportive Use of Adipose-Derived Stem/Stromal Cells , 2007, Aesthetic Plastic Surgery.

[15]  J. Gimble,et al.  Adipose-derived stem cells for regenerative medicine. , 2007, Circulation research.

[16]  K. Yoshimura,et al.  Cell-assisted lipotransfer: supportive use of human adipose-derived cells for soft tissue augmentation with lipoinjection. , 2006, Tissue engineering.

[17]  J. Gimble,et al.  The Immunogenicity of Human Adipose‐Derived Cells: Temporal Changes In Vitro , 2006, Stem cells.

[18]  Sanjin Zvonic,et al.  Immunophenotype of Human Adipose‐Derived Cells: Temporal Changes in Stromal‐Associated and Stem Cell–Associated Markers , 2006, Stem cells.

[19]  J. Rubin,et al.  Adipose tissue: stem cells and beyond. , 2006, Clinics in plastic surgery.

[20]  D. Prockop,et al.  Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. , 2006, Cytotherapy.

[21]  D. García-Olmo,et al.  A Phase I Clinical Trial of the Treatment of Crohn’s Fistula by Adipose Mesenchymal Stem Cell Transplantation , 2005, Diseases of the colon and rectum.

[22]  A. Jödicke,et al.  Autologous stem cells (adipose) and fibrin glue used to treat widespread traumatic calvarial defects: case report. , 2004, Journal of cranio-maxillo-facial surgery : official publication of the European Association for Cranio-Maxillo-Facial Surgery.

[23]  J. Gimble,et al.  Yield of human adipose-derived adult stem cells from liposuction aspirates. , 2004, Cytotherapy.

[24]  D. García-Olmo,et al.  Autologous stem cell transplantation for treatment of rectovaginal fistula in perianal Crohn's disease: a new cell-based therapy , 2003, International Journal of Colorectal Disease.

[25]  F. Guilak,et al.  Adipose-derived adult stem cells: isolation, characterization, and differentiation potential. , 2003, Cytotherapy.

[26]  Min Zhu,et al.  Human adipose tissue is a source of multipotent stem cells. , 2002, Molecular biology of the cell.

[27]  W. Wilkison,et al.  Extracellular matrix mineralization and osteoblast gene expression by human adipose tissue-derived stromal cells. , 2001, Tissue engineering.

[28]  J. Gimble,et al.  Surface protein characterization of human adipose tissue‐derived stromal cells , 2001, Journal of cellular physiology.

[29]  W. Wilkison,et al.  Thiazolidinediones and glucocorticoids synergistically induce differentiation of human adipose tissue stromal cells: biochemical, cellular, and molecular analysis. , 2001, Metabolism: clinical and experimental.

[30]  H. Lorenz,et al.  Multilineage cells from human adipose tissue: implications for cell-based therapies. , 2001, Tissue engineering.