Overcoming the bottleneck of platelet lysate supply in large-scale clinical expansion of adipose-derived stem cells: A comparison of fresh versus three types of platelet lysates from outdated buffy coat–derived platelet concentrates
| Autor: | Rehannah Borup, Anne Fischer-Nielsen, Jens Jørgen Elberg, Mikkel Herly, Jesper Dyrendom Svalgaard, Maj-Lis Møller Talman, Anders Bruun Mathiasen, Krzysztof T. Drzewiecki, Stig-Frederik Trojahn Kølle, Peter V. Glovinski |
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| Rok vydání: | 2017 |
| Předmět: |
Adult
Blood Platelets Cell Extracts Cancer Research Time Factors medicine.medical_treatment Immunology Cell Culture Techniques Cell Separation Platelet Transfusion Buffy coat 030204 cardiovascular system & hematology Biology Andrology Plasma 03 medical and health sciences 0302 clinical medicine Refrigeration Freezing medicine Humans Immunology and Allergy Doubling time Platelet Genetics (clinical) Cell Proliferation Blood Specimen Collection Transplantation Cluster of differentiation Platelet-Rich Plasma Cell growth Growth factor Cell Biology Culture Media Adult Stem Cells Adipose Tissue Oncology Blood Preservation Cell culture 030220 oncology & carcinogenesis Blood Buffy Coat Female Platelet lysate |
| Zdroj: | Cytotherapy. 19:222-234 |
| ISSN: | 1465-3249 |
| DOI: | 10.1016/j.jcyt.2016.10.014 |
| Popis: | Background Platelet lysates (PL) represent a promising replacement for xenogenic growth supplement for adipose-derived stem cell (ASC) expansions. However, fresh platelets from human blood donors are not clinically feasible for large-scale cell expansion based on their limited supply. Therefore, we tested PLs prepared via three methods from outdated buffy coat–derived platelet concentrates (PCs) to establish an efficient and feasible expansion of ASCs for clinical use. Methods PLs were prepared by the freeze-thaw method from freshly drawn platelets or from outdated buffy coat–derived PCs stored in the platelet additive solution, InterSol. Three types of PLs were prepared from outdated PCs with platelets suspended in either (1) InterSol (not manipulated), (2) InterSol + supplemented with plasma or (3) plasma alone (InterSol removed). Using these PLs, we compared ASC population doubling time, cell yield, differentiation potential and cell surface markers. Gene expression profiles were analyzed using microarray assays, and growth factor concentrations in the cell culture medium were measured using enzyme-linked immunosorbent assay (ELISA). Results Of the three PL compositions produced from outdated PCs, removal of Intersol and resuspension in plasma prior to the first freezing process was overall the best. This specific outdated PL induced ASC growth kinetics, surface markers, plastic adherence and differentiation potentials comparable with PL from fresh platelets. ASCs expanded in PL from fresh versus outdated PCs exhibited different expressions of 17 overlapping genes, of which 10 were involved in cellular proliferation, although not significantly reflected by cell growth. Only minor differences in growth factor turnover were observed. Conclusion PLs from outdated platelets may be an efficient and reliable source of human growth supplement allowing for large-scale ASC expansion for clinical use. |
| Databáze: | OpenAIRE |
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