Abstract
Recently, various studies using adipose-derived stem cells (ADSCs) have been performed. However, the safety of ADSCs has not been determined, and protocols for isolating ADSCs have not been established. This study evaluated the activity and toxicity of residual collagenase in isolated ADSCs and the carcinogenicity of these cells. It evaluated the current use of ADSC-related procedures in South Korea as reference data for the authors’ studies. The study surveyed 100 private plastic surgical clinics, 68 plastic surgery departments at general and university hospitals, and 5 biotechnology companies by telephone. Among these, 14 institutions were surveyed using a more detailed questionnaire about ADSC-related procedures and methods of processing adipose tissue. The survey also evaluated the residual collagenase activity during five washes of the ADSC isolation procedure with furyl acryloyl-Leu-Gly-Pro-Ala (FALGPA) and ninhydrin assays. A 4-week toxicity study in non-obese diabetes/severe combined immunodeficiency (NOD/SCID) mice was performed as well as a tumorigenicity study in BALB/c-nu mice using ADSCs from the first and third washes. According to the findings, ADSC-related procedures were performed in 16 % of the private clinics and 14.7 % of the general hospitals surveyed. Among the 14 institutions, 0.1 % type 1 collagenase was used most frequently, and three washes generally were performed. After the first wash, residual collagenase activity was the same as in the blank group (saline only). No toxicity resulting from residual collagenase or tumorigenicity associated with the ADSCs was observed. The results of the current study may be beneficial for establishing safe ADSC isolation protocols and can be used as fundamental data for clinical applications involving ADSCs.
Level of Evidence II
This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.
Similar content being viewed by others
References
Aguiari P, Leo S, Zavan B et al (2008) High glucose induces adipogenic differentiation of muscle-derived stem cells. Proc Natl Acad Sci USA 105:1226–1231
Bunnell BA, Flaat M, Gagliardi C et al (2008) Adipose-derived stem cells: isolation, expansion, and differentiation. Methods 45:115–120
Espey LL, Rondell P (1967) Estimation of mammalian collagenolytic activity with a synthetic substrate. J Appl Physiol 23:757–761
Kahn P, Shin SI (1979) Cellular tumorigenicity in nude mice: test of associations among loss of cell-surface fibronectin, anchorage independence, and tumor-forming ability. J Cell Biol 82:1–16
Kim WS, Park BS, Sung JH (2009) Protective role of adipose-derived stem cells and their soluble factors in photoaging. Arch Dermatol Res 301:329–336
Kim WS, Park BS, Sung JH (2009) The wound-healing and antioxidant effects of adipose-derived stem cells. Expert Opin Biol Ther 9:879–887
Komarova S, Kawakami Y, Stoff-Khalili MA et al (2006) Mesenchymal progenitor cells as cellular vehicles for delivery of oncolytic adenoviruses. Mol Cancer Ther 5:755–766
Kucerova L, Altanerova V, Matuskova M et al (2007) Adipose tissue-derived human mesenchymal stem cells mediated pro-drug cancer gene therapy. Cancer Res 67:6304–6313
Lawrenz B, Schiller H, Willbold E et al (2004) Highly sensitive biosafety model for stem-cell-derived grafts. Cytotherapy 6:212–222
Mandl I, MacLennan JD, Howes EL (1953) Isolation and characterization of proteinase and collagenase from Clostridium histolyticum. J Clin Invest 32:1323–1329
Mizuno H (2009) Adipose-derived stem cells for tissue repair and regeneration: ten years of research and a literature review. J Nippon Med Sch 76:56–66
Moseley TA, Zhu M, Hedrick MH et al (2006) Adipose-derived stem and progenitor cells as fillers in plastic and reconstructive surgery. Plast Reconstr Surg 118:121S–128S
Rodbell M (1967) Metabolism of isolated fat cells: V. Preparation of “ghosts” and their properties: adenyl cyclase and other enzymes. J Biol Chem 242:5744–5750
Rubio D, Garcia-Castro J, Martin MC et al (2005) Spontaneous human adult stem cell transformation. Cancer Res 65:3035–3039
Schaffler A, Buchler C (2007) Concise review: adipose tissue-derived stromal cells: basic and clinical implications for novel cell-based therapies. Stem Cells 25:818–827
van Wart HE, Steinbrink DR (1981) A continuous spectrophotometric assay for Clostridium histolyticum collagenase. Anal Biochem 113:356–365
Wasim SK, Adetola BA, Timothy EH (2007) Hypoxic conditions increase hypoxia-inducible transcription factor 2α and enhance chondrogenesis in stem cells from the infrapatellar fat pad of osteoarthritis patients. Arthritis Res Ther 9:R55
Yoshimura K, Sato K, Aoi N et al (2008) Cell-assisted lipotransfer for cosmetic breast augmentation: supportive use of adipose-derived stem/stromal cells. Aesthetic Plast Surg 32:48–55
Zuk PA, Zuk M, Ashjian P et al (2002) Human adipose tissue is a source for multipotent stem cells. Mol Biol Cell 13:4279–4295
Zuk PA, Zuk M, Mizuno H et al (2001) Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 7:211–228
Acknowledgments
This research was support by a Grant (08122-531) from the Korea Food and Drug Administration in 2008.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Chang, H., Do, BR., Che, JH. et al. Safety of Adipose-Derived Stem Cells and Collagenase in Fat Tissue Preparation. Aesth Plast Surg 37, 802–808 (2013). https://doi.org/10.1007/s00266-013-0156-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00266-013-0156-7