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Quality Control Platform for the Standardization of a Regenerative Medicine Product

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...

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Autores principales: Zia, Silvia, Roda, Barbara, Zannini, Chiara, Alviano, Francesco, Bonsi, Laura, Govoni, Marco, Vivarelli, Leonardo, Fazio, Nicola, Dallari, Dante, Reschiglian, Pierluigi, Zattoni, Andrea
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9026409/
https://www.ncbi.nlm.nih.gov/pubmed/35447702
http://dx.doi.org/10.3390/bioengineering9040142
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author Zia, Silvia
Roda, Barbara
Zannini, Chiara
Alviano, Francesco
Bonsi, Laura
Govoni, Marco
Vivarelli, Leonardo
Fazio, Nicola
Dallari, Dante
Reschiglian, Pierluigi
Zattoni, Andrea
author_facet Zia, Silvia
Roda, Barbara
Zannini, Chiara
Alviano, Francesco
Bonsi, Laura
Govoni, Marco
Vivarelli, Leonardo
Fazio, Nicola
Dallari, Dante
Reschiglian, Pierluigi
Zattoni, Andrea
author_sort Zia, Silvia
collection PubMed
description 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.
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spelling pubmed-90264092022-04-23 Quality Control Platform for the Standardization of a Regenerative Medicine Product Zia, Silvia Roda, Barbara Zannini, Chiara Alviano, Francesco Bonsi, Laura Govoni, Marco Vivarelli, Leonardo Fazio, Nicola Dallari, Dante Reschiglian, Pierluigi Zattoni, Andrea Bioengineering (Basel) Article 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. MDPI 2022-03-28 /pmc/articles/PMC9026409/ /pubmed/35447702 http://dx.doi.org/10.3390/bioengineering9040142 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zia, Silvia
Roda, Barbara
Zannini, Chiara
Alviano, Francesco
Bonsi, Laura
Govoni, Marco
Vivarelli, Leonardo
Fazio, Nicola
Dallari, Dante
Reschiglian, Pierluigi
Zattoni, Andrea
Quality Control Platform for the Standardization of a Regenerative Medicine Product
title Quality Control Platform for the Standardization of a Regenerative Medicine Product
title_full Quality Control Platform for the Standardization of a Regenerative Medicine Product
title_fullStr Quality Control Platform for the Standardization of a Regenerative Medicine Product
title_full_unstemmed Quality Control Platform for the Standardization of a Regenerative Medicine Product
title_short Quality Control Platform for the Standardization of a Regenerative Medicine Product
title_sort quality control platform for the standardization of a regenerative medicine product
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9026409/
https://www.ncbi.nlm.nih.gov/pubmed/35447702
http://dx.doi.org/10.3390/bioengineering9040142
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