A comprehensive characterisation of large-scale expanded human bone marrow and umbilical cord mesenchymal stem cells

Autor: Karina T. Wright, Charlotte H Hulme, Sally Roberts, John Garcia, Claire Mennan
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Adult
Male
Pathology
medicine.medical_specialty
Multipotential differentiation
Stromal cell
Cell Culture Techniques
Medicine (miscellaneous)
Bone Marrow Cells
Cell Separation
Biology
Q1
Biochemistry
Genetics and Molecular Biology (miscellaneous)
Umbilical cord
Flow cytometry
Large-scale expansion
lcsh:Biochemistry
03 medical and health sciences
Tissue culture
0302 clinical medicine
medicine
Humans
Bone marrow
lcsh:QD415-436
030304 developmental biology
Hollow fibre bioreactor
0303 health sciences
lcsh:R5-920
Telomere length
medicine.diagnostic_test
Cluster of differentiation
Characterisation
Research
Mesenchymal stem cell
Infant
Newborn
Cell Differentiation
Cell Biology
R1
medicine.anatomical_structure
030220 oncology & carcinogenesis
Molecular Medicine
Mesenchymal stem cells
Stem cell
lcsh:Medicine (General)
Zdroj: Stem Cell Research & Therapy, Vol 10, Iss 1, Pp 1-15 (2019)
Stem Cell Research & Therapy
ISSN: 1757-6512
DOI: 10.1186/s13287-019-1202-4
Popis: Background The manufacture of mesenchymal stem/stromal cells (MSCs) for clinical use needs to be cost effective, safe and scaled up. Current methods of expansion on tissue culture plastic are labour-intensive and involve several ‘open’ procedures. We have used the closed Quantum® hollow fibre bioreactor to expand four cultures each of MSCs derived from bone marrow (BM) and, for the first time, umbilical cords (UCs) and assessed extensive characterisation profiles for each, compared to parallel cultures grown on tissue culture plastic. Methods Bone marrow aspirate was directly loaded into the Quantum®, and cells were harvested and characterised at passage (P) 0. Bone marrow cells were re-seeded into the Quantum®, harvested and further characterised at P1. UC-MSCs were isolated enzymatically and cultured once on tissue culture plastic, before loading cells into the Quantum®, harvesting and characterising at P1. Quantum®-derived cultures were phenotyped in terms of immunoprofile, tri-lineage differentiation, response to inflammatory stimulus and telomere length, as were parallel cultures expanded on tissue culture plastic. Results Bone marrow cell harvests from the Quantum® were 23.1 ± 16.2 × 106 in 14 ± 2 days (P0) and 131 ± 84 × 106 BM-MSCs in 13 ± 1 days (P1), whereas UC-MSC harvests from the Quantum® were 168 ± 52 × 106 UC-MSCs after 7 ± 2 days (P1). Quantum®- and tissue culture plastic-expanded cultures at P1 adhered to criteria for MSCs in terms of cell surface markers, multipotency and plastic adherence, whereas the integrins, CD29, CD49c and CD51/61, were found to be elevated on Quantum®-expanded BM-MSCs. Rapid culture expansion in the Quantum® did not cause shortened telomeres when compared to cultures on tissue culture plastic. Immunomodulatory gene expression was variable between donors but showed that all MSCs upregulated indoleamine 2, 3-dioxygenase (IDO). Conclusions The results presented here demonstrate that the Quantum® can be used to expand large numbers of MSCs from bone marrow and umbilical cord tissues for next-generation large-scale manufacturing, without impacting on many of the properties that are characteristic of MSCs or potentially therapeutic. Using the Quantum®, we can obtain multiple MSC doses from a single manufacturing run to treat many patients. Together, our findings support the development of cheaper cell-based treatments. Electronic supplementary material The online version of this article (10.1186/s13287-019-1202-4) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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