Bioimpacts. 2019;9(3): 161-172.
doi: 10.15171/bi.2019.20
PMID: 31508331
PMCID: PMC6726751
Scopus ID: 85071165666
WOSID: 000483926500005
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Original Research

A short-term plastic adherence incubation of the stromal vascular fraction leads to a predictable GMP-compliant cell-product

Stephan Born 1 * , Max Johannes Dörfel 1, Philip Hartjen 2, Seyed Ali Haschemi Yekani 1, Julia Luecke 1, Juliane Katharina Meutsch 1, Julie Katharina Westphal 1, Moritz Birkelbach 2, Robert Köhnke 2, Ralf Smeets 2,3, Michael Krueger 1

1 OXACELL® AG, Potsdam, Germany
2 Department of Oral and Maxillofacial Surgery, University Hospital Hamburg-Eppendorf, Hamburg, Germany
3 Division, Regenerative Orofacial Medicine, University Hospital Hamburg-Eppendorf, Hamburg, Germany


Introduction: Mesenchymal stromal/stem cells (MSCs) derived from fat tissue are an encouraging tool for regenerative medicine. They share properties similar to the bone marrow-derived MSCs, but the amount of MSCs per gram of fat tissue is 500x higher. The fat tissue can easily be digested by collagenase, releasing a heterogeneous cell fraction called stromal vascular fraction (SVF) which contains a variable amount of stromal/stem cells. In Europe, cell products like the SVF derived from fat tissue are considered advanced therapy medicinal product (ATMPs). As a consequence, the manufacturing process has to be approved via GMP-compliant process validation. The problem of the process validation for SVF is the heterogeneity of this fraction.
Methods: Here, we modified existing purification strategies by adding an additional plastic adherence incubation of maximal 20 hours after SVF isolation. The resulting cell fraction was characterized and compared to SVF as well as cultivated adipose-derived stromal/stem cells (ASCs) with respect to viability and cell yield, the expression of surface markers, differentiation potential and cytokine expression.
Results: Short-term incubation significantly reduced the heterogeneity of the resulting cell fraction compared to SVF. The cells were able to differentiate into adipocytes, chondrocytes, and osteoblasts. More importantly, they expressed trophic proteins which have been previously associated with the beneficial effects of MSCs. Furthermore, GMP compliance of the production process described herein was acknowledged by the national regulatory agencies (DE_BB_01_GMP_2017_1018).
Conclusion: Addition of a short purification-step after the SVF isolation is a cheap and fast strategy to isolate a homogeneous uncultivated GMP-compliant cell fraction of ASCs.
Keywords: Adipose derived stromal/stem cells, Stromal vascular fraction, Mesenchymal stromal/stem cells, Cell therapy, Regenerative medicine, Good manufacturing practice
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Submitted: 14 Jan 2019
Revision: 07 Mar 2019
Accepted: 08 Mar 2019
ePublished: 25 Mar 2019
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