A standardized procedure to obtain mesenchymal stem/stromal cells from minimally manipulated dental pulp and Wharton’s jelly samples

Authors

  • Maxime Ducret Laboratoire de Biologie Tissulaire et Ingénierie thérapeutique, UMR5305 CNRS/Université Lyon 1, Institut de Biologie et Chimie des Protéines, 7 passage du Vercors, 69367 Lyon Cedex 07, France
  • H. Fabre 1Laboratoire de Biologie Tissulaire et Ingénierie thérapeutique, UMR5305 CNRS/Université Lyon 1, Institut de Biologie et Chimie des Protéines, 7 passage du Vercors, 69367 Lyon Cedex 07, France
  • O. Degoult BIOTECH, Cell Therapy Research Institute, 5 avenue Lionel Terray, 69330 Meyzieu, France
  • G. Atzeni BIOTECH, Cell Therapy Research Institute, 5 avenue Lionel Terray, 69330 Meyzieu, France
  • C. McGuckin BIOTECH, Cell Therapy Research Institute, 5 avenue Lionel Terray, 69330 Meyzieu, France
  • N. Forraz BIOTECH, Cell Therapy Research Institute, 5 avenue Lionel Terray, 69330 Meyzieu, France
  • F. Mallein-Gerin 1Laboratoire de Biologie Tissulaire et Ingénierie thérapeutique, UMR5305 CNRS/Université Lyon 1, Institut de Biologie et Chimie des Protéines, 7 passage du Vercors, 69367 Lyon Cedex 07, France
  • E. Perrier-Groult Laboratoire de Biologie Tissulaire et Ingénierie thérapeutique, UMR5305 CNRS/Université Lyon 1, Institut de Biologie et Chimie des Protéines, 7 passage du Vercors, 69367 Lyon Cedex 07, France
  • J.C. Farges 1Laboratoire de Biologie Tissulaire et Ingénierie thérapeutique, UMR5305 CNRS/Université Lyon 1, Institut de Biologie et Chimie des Protéines, 7 passage du Vercors, 69367 Lyon Cedex 07, France 2Université de Lyon, Université Lyon 1, Faculté d’Odontologie, 11 rue Guillaume Paradin, 69372 Lyon Cedex 08, France 3Hospices Civils de Lyon, Service de Consultations et Traitements Dentaires, 6-8 place Depéret, 69007 Lyon, France

Keywords:

Human dental pulp, Wharton’s jelly, stem/ stromal cells, good manufacturing practices, cell-based medicinal products, immunophenotyping

Abstract

Transplantation of mesenchymal stem/stromal cells (MSCs) has emerged as an effective method to treat diseased or damaged organs and tissues, and hundreds of clinical trials using MSCs are currently under way to demonstrate the validity of such a therapeutic approach. However, most MSCs used for clinical trials are prepared in research laboratories with insufficient manufacturing quality control.
In particular, laboratories lack standardized procedures for in vitro isolation of MSCs from tissue samples, resulting in heterogeneous populations of cells and variable experimental and clinical results. MSCs are now referred to as Human Cellular Tissue-based Products or Advanced Therapy Medicinal Products, and guidelines from the American Code of Federal Regulation of the Food and Drug Administration (21 CFR Part 1271) and from the European Medicines Agency (European Directive 1394/2007) define requirements for appropriate production of these cells. These guidelines, commonly called “Good Manufacturing Practices” (GMP), include recommendations about laboratory cell culture procedures to ensure optimal reproducibility, efficacy and safety of the final medicinal product. In particular, the Food and Drug Administration divides ex vivo cultured cells into “minimally” and “more than minimally” manipulated samples, in function of the use or not of procedures “that might alter the biological features of the cells”. Today, minimal manipulation conditions have not been defined for the collection and isolation of MSCs (Torre et al. 2015)(Ducret et al. 2015).
Most if not all culture protocols that have been reported so far are unsatisfactory, because of the use of xeno- or allogeneic cell culture media, enzymatic treatment and long-term cell amplification that are known to alter the quality of MSCs. The aim of this study was to describe a standardized procedure for recovering MSCs with minimal handling from two promising sources, the dental pulp (DP) and the Wharton’s jelly (WJ) of the umbilical cord. The quality and homogeneity of the expanded cell populations were assessed by using flow cytometry with criteria that go beyond the International Society of Cellular Therapy (ISCT) guidelines for MSC characterization.

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Published

2016-06-28

Issue

Vol. 53 No. 1 (2016)

Section

Articles

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