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Quality Control Platform for the Standardization of a Regenerative Medicine Product

Authors :
Silvia Zia
Barbara Roda
Chiara Zannini
Francesco Alviano
Laura Bonsi
Marco Govoni
Leonardo Vivarelli
Nicola Fazio
Dante Dallari
Pierluigi Reschiglian
Andrea Zattoni
Source :
Bioengineering, Vol 9, Iss 4, p 142 (2022)
Publication Year :
2022
Publisher :
MDPI AG, 2022.

Abstract

Adipose tissue is an attractive source of stem cells due to its wide availability. They contribute to the stromal vascular fraction (SVF), which is composed of pre-adipocytes, tissue-progenitors, and pericytes, among others. Because its direct use in medical applications is increasing worldwide, new quality control systems are required. We investigated the ability of the Non-Equilibrium Earth Gravity Assisted Dynamic Fractionation (NEEGA-DF) method to analyze and separate cells based solely on their physical characteristics, resulting in a fingerprint of the biological sample. Adipose tissue was enzymatically digested, and the SVF was analyzed by NEEGA-DF. Based on the fractogram (the UV signal of eluting cells versus time of analysis) the collection time was set to sort alive cells. The collected cells (F-SVF) were analyzed for their phenotype, immunomodulation ability, and differentiation potential. The SVF profile showed reproducibility, and the alive cells were collected. The F-SVF showed intact adhesion phenotype, proliferation, and differentiation potential. The methodology allowed enrichment of the mesenchymal component with a higher expression of mesenchymal markers and depletion of debris, RBCs, and an extracellular matrix still present in the digestive product. Moreover, cells eluting in the last minutes showed higher circularity and lower area, proving the principles of enrichment of a more homogenous cell population with better characteristics. We proved the NEEGA-DF method is a “gentle” cell sorter that purifies primary cells obtained by enzymatic digestion and does not alter any stem cell function.

Details

Language :
English
ISSN :
23065354
Volume :
9
Issue :
4
Database :
Directory of Open Access Journals
Journal :
Bioengineering
Publication Type :
Academic Journal
Accession number :
edsdoj.385a27d03600468f9fec338affbfb931
Document Type :
article
Full Text :
https://doi.org/10.3390/bioengineering9040142