A New Human Platelet Lysate for Mesenchymal Stem Cell Production Compliant with Good Manufacturing Practice Conditions
| Autor: | Katia Mareschi, Elena Marini, Alessia Giovanna Santa Banche Niclot, Marta Barone, Giuseppe Pinnetta, Aloe Adamini, Manuela Spadea, Luciana Labanca, Graziella Lucania, Ivana Ferrero, Franca Fagioli |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2022 |
| Předmět: |
Blood Platelets
Organic Chemistry Cell Differentiation Mesenchymal Stem Cells General Medicine Catalysis Culture Media Immunophenotyping Computer Science Applications Inorganic Chemistry Humans Intercellular Signaling Peptides and Proteins Physical and Theoretical Chemistry Molecular Biology Cells Cultured mesenchymal stem cells human platelet lysate GMP Spectroscopy Cell Proliferation |
| Zdroj: | International Journal of Molecular Sciences; Volume 23; Issue 6; Pages: 3234 |
| Popis: | Mesenchymal stem cells (MSCs) are classified as advanced therapy medicinal products, a new category of GMP (good manufacturing practice)-compliant medicines for clinical use. We isolated MSCs from 5 bone marrow (BM) samples using human platelet lysate (HPL) instead of foetal bovine serum (FBS). We used a new method of HPL production consisting of treating platelet (PLTs) pools with Ca-Gluconate to form a gel clot, then mechanically squeezing to release growth factors. We compared the new HPL (HPL-S) with the standard (HPL-E) obtained by freezing/thawing cycles and by adding heparin. HPL-S had not PLTs and fibrinogen but the quantity of proteins and growth factors was comparable to HPL-E. Therefore, HPL-S needed fewer production steps to be in compliance with GMP conditions. The number of colonies forming unit-fibroblasts (CFU-F) and the maintenance of stem markers showed no significant differences between MSCs with HPL-E and HPL-S. The cumulative population doubling was higher in MSCs with HPL-E in the earlier passages, but we observed an inverted trend of cell growth at the fourth passage. Immunophenotypic analysis showed a significant lower expression of HLA-DR in the MSCs with HPL-S (1.30%) than HPL-E (14.10%). In conclusion, we demonstrated that HPL-S is an effective alternative for MSC production under GMP conditions. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|