Large-Scale Automated Hollow-Fiber Bioreactor Expansion of Umbilical Cord-Derived Human Mesenchymal Stromal Cells for Neurological Disorders
Autor: | Tereza Kučírková, Hana Lejdarova, Veronika Oralova, Eva Matalová, Serhiy Forostyak, Petr Kuglík, Zdenek Koristek, Veronika Pospisilova, Ladislava Vymetalova, Eva Makaturova, Lucia Knopfová, Tomas Kasko, Petr Beneš |
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Rok vydání: | 2019 |
Předmět: |
0301 basic medicine
endocrine system medicine.medical_treatment Biology Mesenchymal Stem Cell Transplantation Biochemistry Regenerative medicine Umbilical cord Umbilical Cord 03 medical and health sciences Cellular and Molecular Neuroscience Paracrine signalling Tissue culture Bioreactors 0302 clinical medicine medicine Humans Wharton Jelly Cells Cultured Cell Proliferation Growth factor Mesenchymal stem cell Mesenchymal Stem Cells General Medicine equipment and supplies medicine.disease Astrogliosis Cell biology 030104 developmental biology medicine.anatomical_structure Nervous System Diseases 030217 neurology & neurosurgery Explant culture |
Zdroj: | Neurochemical Research. 45:204-214 |
ISSN: | 1573-6903 0364-3190 |
Popis: | Neurodegenerative disorders present a broad group of neurological diseases and remain one of the greatest challenges and burdens to mankind. Maladies like amyotrophic lateral sclerosis, Alzheimer's disease, stroke or spinal cord injury commonly features astroglia involvement (astrogliosis) with signs of inflammation. Regenerative, paracrine and immunomodulatory properties of human mesenchymal stromal cells (hMSCs) could target the above components, thus opening new therapeutic possibilities for regenerative medicine. A special interest should be given to hMSCs derived from the umbilical cord (UC) tissue, due to their origin, properties and lack of ethical paradigms. The aim of this study was to establish standard operating and scale-up good manufacturing practice (GMP) protocols of UC-hMSCs isolation, characterization, expansion and comparison of cells' properties when harvested on T-flasks versus using a large-scale bioreactor system. Human UC-hMSCs, isolated by tissue explant culture technique from Wharton's jelly, were harvested after reaching 75% confluence and cultured using tissue culture flasks. Obtained UC-hMSCs prior/after the cryopreservation and after harvesting in a bioreactor, were fully characterized for "mesenchymness" immunomodulatory, tumorigenicity and genetic stability, senescence and cell-doubling properties, as well as gene expression features. Our study demonstrates an efficient and simple technique for large scale UC-hMSCs expansion. Harvesting of UC-hMSCs' using classic and large scale methods did not alter UC-hMSCs' senescence, genetic stability or in vitro tumorigenicity features. We observed comparable growth and immunomodulatory capacities of fresh, frozen and expanded UC-hMSCs. We found no difference in the ability to differentiate toward adipogenic, osteogenic and chondrogenic lineages between classic and large scale UC-hMSCs expansion methods. Both, methods enabled derivation of genetically stabile cells with typical mesenchymal features. Interestingly, we found significantly increased mRNA expression levels of neural growth factor (NGF) and downregulated insulin growth factor (IGF) in UC-hMSCs cultured in bioreactor, while IL4, IL6, IL8, TGFb and VEGF expression levels remained at the similar levels. A culturing of UC-hMSCs using a large-scale automated closed bioreactor expansion system under the GMP conditions does not alter basic "mesenchymal" features and quality of the cells. Our study has been designed to pave a road toward translation of basic research data known about human UC-MSCs for the future clinical testing in patients with neurological and immunocompromised disorders. An industrial manufacturing of UC-hMSCs next will undergo regulatory approval following advanced therapy medicinal products (ATMP) criteria prior to clinical application and approval to be used in patients. |
Databáze: | OpenAIRE |
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