Bioimpacts. 2019;9(3):161-167.
doi: 10.15171/bi.2019.20
  Abstract View: 105
  PDF Download: 2

Original Research

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

Abstract

Introduction

Mesenchymal stromal/stem cells derived from fat tissue are an encouraging tool for regenerative medicine. They share properties similar to bone marrow derived mesenchymal stromal/stem cells but the amount of mesenchymal stromal/stem cells 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 Products (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 20h after SVF isolation. The resulting cell fraction was characterized and compared to SVF as well as cultivated adipose-derived stromal/stem cells with respect to viability and cell yield, the expression of surface markers, differentiation potential and cytokine expression.

Results

Short-term incubation significantly reduced 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.

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Submitted: 14 Jan 2019
Revised: 07 Mar 2019
Accepted: 08 Mar 2019
First published online: 08 Mar 2019
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