Minimally manipulated whole human umbilical cord is a rich source of clinical-grade human mesenchymal stromal cells expanded in human platelet lysate

Abstract Background aims Mesenchymal stromal cells (MSC) have recently been identified as a therapeutic option in several clinical conditions. Whereas bone marrow (BM) is considered the main source of MSC (BM-MSC), the invasive technique required for collection and the decline in allogeneic donation...

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Published in:Cytotherapy (Oxford, England) England), 2011-08, Vol.13 (7), p.786-801
Main Authors: Capelli, Chiara, Gotti, Elisa, Morigi, Marina, Rota, Cinzia, Weng, Ling, Dazzi, Francesco, Spinelli, Orietta, Cazzaniga, Giovanni, Trezzi, Rosangela, Gianatti, Andrea, Rambaldi, Alessandro, Golay, Josee, Introna, Martino
Format: Article
Language:English
Subjects:
Adipogenesis
Advanced Basic Science
Animals
Blood Platelets - cytology
Carcinogenicity Tests
Cell Differentiation
Cell Lineage
Cells, Cultured
Comparative Genomic Hybridization
Culture Media
Disease Models, Animal
Fibroblasts - cytology
good manufacturing practice
Graft vs Host Disease - immunology
Humans
Immunosuppressive Agents - pharmacology
Mesenchymal Stem Cells - immunology
mesenchymal stromal cells
Mice
Mice, Inbred BALB C
Mice, SCID
Other
platelet lysate
Stem Cells
umbilical cord
Umbilical Cord - cytology
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Summary:Abstract Background aims Mesenchymal stromal cells (MSC) have recently been identified as a therapeutic option in several clinical conditions. Whereas bone marrow (BM) is considered the main source of MSC (BM-MSC), the invasive technique required for collection and the decline in allogeneic donations call for alternative sources. Human umbilical cord (UC) represents an easily available source of MSC (UC-MSC). Methods Sections of full-term UC were transferred to cell culture flasks and cultured in 5% human platelet lysate (PL)-enriched medium. Neither enzymatic digestion nor blood vessel removal was performed. After 2 weeks, the adherent cells were harvested (P1), replated at low density and expanded for two consecutive rounds (P2 and P3). Results We isolated and expanded MSC from 9/9 UC. UC-MSC expanded with a mean fold increase (FI) of 42 735 ± 16 195 from P1 to P3 in a mean of 29 ± 2 days. By processing the entire cord unit, we theoretically could have reached a median of 9.5 × 1010 cells (ranging from 1.0 × 1010 to 29.0 × 1010 ). UC-MSC expressed standard surface markers; they contained more colony-forming unit (CFU)-fibroblast (F) and seemed less committed towards osteogenic, chondrogenic and adipogenic lineages than BM-MSC. They showed immunosuppressive properties both in vitro and in an in vivo chronic Graft versus Host disease (cGvHD) mouse model. Both array-Comparative Genomic Hybridization (CGH) analysis and karyotyping revealed no chromosome alterations at the end of the expansion. Animal studies revealed no tumorigenicity in vivo. Conclusions UC constitute a convenient and very rich source of MSC for the production of third-party ‘clinical doses’ of cells under good manufacturing practice (GMP) conditions.
ISSN:1465-3249
1477-2566
DOI:10.3109/14653249.2011.563294