The Utilization of Freezing Steps in Mesenchymal Stromal Cell (MSC) Manufacturing: Potential Impact on Quality and Cell Functionality Attributes

Frontiers in Immunology
Sofia OjaJohanna Nystedt

Abstract

Some recent reports suggest that cryopreserved and thawed mesenchymal stromal cells (MSCs) may have impaired functional properties as compared to freshly harvested MSCs from continuous cultures. A cryopreservation step in the manufacturing process brings important benefits, since it enables immediate off-the-shelf access to the products and a completion of all quality testing before batch release and administration to the patient. Cryopreservation is also inevitable in MSC banking strategies. In this study, we present the results from the MSC stability testing program of our in-house manufactured clinical-grade allogeneic bone marrow-derived MSC product that is expanded in platelet lysate and frozen in passage 2. The current manufacturing protocol contains only one freezing step and the frozen MSC product is thawed bed-side at the clinic. We can conclude superior viability and cell recovery of the frozen and thawed MSC product utilizing the validated freezing and thawing protocols we have developed. The MSC phenotype and differentiation potential was generally found to be unaltered after thawing, but the thawed cells exhibited a 50% reduced, but not completely abolished, performance in an in vitro immunosuppression assay. The i...Continue Reading

References

May 13, 2008·Lancet·Katarina Le BlancUNKNOWN Developmental Committee of the European Group for Blood and Marrow Transplantation
Jun 23, 2009·Biology of Blood and Marrow Transplantation : Journal of the American Society for Blood and Marrow Transplantation·Partow KebriaeiJoseph Uberti
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Dec 11, 2013·Biology of Blood and Marrow Transplantation : Journal of the American Society for Blood and Marrow Transplantation·Martino IntronaEttore Biagi
Feb 25, 2014·Nature Biotechnology·James A AnkrumJeffrey M Karp
Jan 8, 2015·Current Opinion in Organ Transplantation·Martino Introna, Alessandro Rambaldi
Feb 26, 2016·Stem Cells and Development·Martin J HoogduijnCarla C Baan
May 22, 2016·Journal of Translational Medicine·Chantal LechanteurYves Beguin
Dec 13, 2016·Bone Marrow Transplantation·U SalmenniemiM Korhonen
Nov 17, 2017·Science Translational Medicine·Antonio GalleuFrancesco Dazzi
Mar 9, 2018·Molecular Therapy. Methods & Clinical Development·Joni KetoJohanna Nystedt
Jun 3, 2018·Cell Stem Cell·Jacques Galipeau, Luc Sensébé

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Citations

Sep 19, 2020·Current Stem Cell Reports·Raghavan ChinnaduraiChristian M Capitini
Nov 25, 2020·Blood Advances·Andre J BurnhamEdwin M Horwitz

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Methods Mentioned

BETA
bone marrow aspiration
density gradient centrifugation
flow cytomety
flow cytometry
FACS
FCS
Assay

Software Mentioned

GraphPad Prism
FlowJo
Cell Insight
GraphPad

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