Validation of an Automated Procedure to Isolate Human Adipose Tissue–Derived Cells by Using the Sepax^®Technology

Abstract

The stromal vascular fraction of adipose tissue has gained popularity as a source of autologous progenitor cells for tissue engineering and regenerative medicine applications. The aim of this study was to validate a newly developed, automated procedure to isolate adipose-derived mesenchymal stem/stromal cells (ASCs) from adult human lipoaspirates in a closed and clinical-grade device, based on the Sepax Ò technology. Using a total of 11 donors, this procedure was compared with the standard operator-based manual separation in terms of isolation yield, clonogenic fraction, phenotype, and differentiation potential of ASCs. As compared with the manual process, automation resulted in a 62% higher isolation yield, with 2.6 -1.2 · 10 5 nucleated cells per mL of liposuction, and a 24% higher frequency of clonogenic progenitors. The variability in the isolation yield and clonogenicity across different preparations was reduced by 18% and 50%, respectively. The cytofluorimetric profile and in vitro differentiation capacity into mesenchymal lineages were comparable in the cells isolated using the two procedures. The new Sepax-based process thus allows an efficient isolation of ASCs with higher and more reproducible yields than the standard manual procedure, along with minimal operator intervention. These results are expected to facilitate the use of ASCs for clinical purposes, either within an intraoperative setting or in combination with further in vitro cell expansion/cultivation.

The Celution®System: Automated Processing of Adipose-Derived Regenerative Cells in a Functionally Closed System

“…However, one recent publication has compared published data obtained with the Celution System 23 with the output of tissue that had been digested using a manual process and then washed and concentrated using the SepaxÔ system (Biosafe SA, Geneva, Switzerland), a product originally designed for processing umbilical cord blood. 31 The results suggest comparability of output, within the limitations of the nonhead-to-head study design. However, with this approach, the digestion component of processing is neither automated nor standardized.…”

Allogeneic and Xenogeneic Transplantation of Adipose-Derived Stem Cells in Immunocompetent Recipients Without Immunosuppressants

“…The SVF can be directly used for therapy or further processed for the isolation of ADSC. In future clinical applications, adipose tissue mincing will undoubtedly be substituted by liposuction, and the SVF isolation handled by all-in-one devices that are now commercially available [5,27,28]. The adoption of these automation procedures has resulted in the high availability of human ADSCs, which could perhaps explain why many preclinical studies chose human ADSCs for transplantation in nonhuman animals such as rats and mice, thus, intentionally or unintentionally demonstrating ADSC's xenotransplantation potential (Tables 1 and 2).…”

“…Liposuction is a well-established clinical practice in human medicine, and processing the liposuction material into SVF can be done expeditiously with commercially available devices [5,27,28]. Therefore, the future application of ADSCs in human medicine will be conducted mostly, if not exclusively, in an autologous fashion.…”

Age-Related Effects on the Potency of Human Adipose-Derived Stem Cells: Creation and Evaluation of Superlots and Implications for Musculoskeletal Tissue Engineering Applications

“…[1][2][3][4][5] Despite their multilineage differentiation potential, hASC exhibit a high level of donor-to-donor variability. [6][7][8][9] This particular characteristic presents a challenge for studying and manipulating hASC on the bench top, as well as employing their widespread use in regenerative medicine applications in the clinic.…”