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

Maxime Ducret, H. Fabre, O. Degoult, G. Atzeni, C. McGuckin, N. Forraz, F. Mallein-Gerin, E. Perrier-Groult, J.C. Farges

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.

Keywords


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

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