Large-Scale Automated Hollow-Fiber Bioreactor Expansion of
Umbilical Cord-Derived Human Mesenchymal Stromal Cells for
Neurological Disorders

J 2020

Large-Scale Automated Hollow-Fiber Bioreactor Expansion of Umbilical Cord-Derived Human Mesenchymal Stromal Cells for Neurological Disorders

VYMETALOVA, Ladislava; Tereza KUCIRKOVA; Lucia KNOPFOVA; Veronika POSPISILOVA; Tomas KASKO et al.

Základní údaje

Originální název

Large-Scale Automated Hollow-Fiber Bioreactor Expansion of Umbilical Cord-Derived Human Mesenchymal Stromal Cells for Neurological Disorders

Autoři

Vydání

Neurochemical Research, New York, Springer, 2020, 0364-3190

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 3.996

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/20:00117084

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

Mesenchymal stromal cells; Umbilical cord tissue; Bioreactor; Large-scale expansion; Good manufacturing practice (GMP)

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 25. 11. 2020 13:27, Mgr. Marie Novosadová Šípková, DiS.

Anotace

V originále

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.

Citovat

VYMETALOVA, Ladislava; Tereza KUCIRKOVA; Lucia KNOPFOVA; Veronika POSPISILOVA; Tomas KASKO; Hana LEJDAROVA; Eva MAKATUROVA; Petr KUGLÍK; Veronika ORALOVA; Eva MATALOVA; Petr BENES; Zdenek KORISTEK a Serhiy FOROSTYAK. Large-Scale Automated Hollow-Fiber Bioreactor Expansion of Umbilical Cord-Derived Human Mesenchymal Stromal Cells for Neurological Disorders. Neurochemical Research. New York: Springer, 2020, roč. 45, č. 1, s. 204-214. ISSN 0364-3190. Dostupné z: https://doi.org/10.1007/s11064-019-02925-y.

@article{1699376,
   author = {Vymetalova, Ladislava and Kucirkova, Tereza and Knopfova, Lucia and Pospisilova, Veronika and Kasko, Tomas and Lejdarova, Hana and Makaturova, Eva and Kuglík, Petr and Oralova, Veronika and Matalova, Eva and Benes, Petr and Koristek, Zdenek and Forostyak, Serhiy},
   article_location = {New York},
   article_number = {1},
   doi = {https://doi.org/10.1007/s11064-019-02925-y},
   keywords = {Mesenchymal stromal cells; Umbilical cord tissue; Bioreactor; Large-scale expansion; Good manufacturing practice (GMP)},
   language = {eng},
   issn = {0364-3190},
   journal = {Neurochemical Research},
   title = {Large-Scale Automated Hollow-Fiber Bioreactor Expansion of Umbilical Cord-Derived Human Mesenchymal Stromal Cells for Neurological Disorders},
   url = {https://doi.org/10.1007/s11064-019-02925-y},
   volume = {45},
   year = {2020}
}

TY  - JOUR
ID  - 1699376
AU  - Vymetalova, Ladislava - Kucirkova, Tereza - Knopfova, Lucia - Pospisilova, Veronika - Kasko, Tomas - Lejdarova, Hana - Makaturova, Eva - Kuglík, Petr - Oralova, Veronika - Matalova, Eva - Benes, Petr - Koristek, Zdenek - Forostyak, Serhiy
PY  - 2020
TI  - Large-Scale Automated Hollow-Fiber Bioreactor Expansion of Umbilical Cord-Derived Human Mesenchymal Stromal Cells for Neurological Disorders
JF  - Neurochemical Research
VL  - 45
IS  - 1
SP  - 204-214
EP  - 204-214
PB  - Springer
SN  - 03643190
KW  - Mesenchymal stromal cells
KW  - Umbilical cord tissue
KW  - Bioreactor
KW  - Large-scale expansion
KW  - Good manufacturing practice (GMP)
UR  - https://doi.org/10.1007/s11064-019-02925-y
L2  - https://doi.org/10.1007/s11064-019-02925-y
N2  - 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.
ER  -

VYMETALOVA, Ladislava; Tereza KUCIRKOVA; Lucia KNOPFOVA; Veronika POSPISILOVA; Tomas KASKO; Hana LEJDAROVA; Eva MAKATUROVA; Petr KUGLÍK; Veronika ORALOVA; Eva MATALOVA; Petr BENES; Zdenek KORISTEK a Serhiy FOROSTYAK. Large-Scale Automated Hollow-Fiber Bioreactor Expansion of Umbilical Cord-Derived Human Mesenchymal Stromal Cells for Neurological Disorders. \textit{Neurochemical Research}. New York: Springer, 2020, roč.~45, č.~1, s.~204-214. ISSN~0364-3190. Dostupné z: https://doi.org/10.1007/s11064-019-02925-y.

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