open access

Vol 54, No 2 (2016)
Original paper
Submitted: 2015-11-19
Accepted: 2016-05-23
Published online: 2016-06-07
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In vitro culture of primary human myoblasts by using the dextran microcarriers Cytodex3®

Natalia Rozwadowska, Agnieszka Malcher, Ewa Baumann, Tomasz Jan Kolanowski, Marek Rucinski, Tomasz Mietkiewski, Katarzyna Fiedorowicz, Maciej Kurpisz
DOI: 10.5603/FHC.a2016.0010
·
Pubmed: 27270505
·
Folia Histochem Cytobiol 2016;54(2):81-90.

open access

Vol 54, No 2 (2016)
ORIGINAL PAPERS
Submitted: 2015-11-19
Accepted: 2016-05-23
Published online: 2016-06-07

Abstract

Introduction. Primary cells in vitro culture scale-up is a crucial issue in cell-based tissue and organ regeneration therapy. Reducing costs and space occupied by the cells cultured in vitro has been an important target. Cells cultured in vitro with the use of bioreactor with dextran microcarriers (Cytodex®) have potentially a chance to meet many of the cell therapy requirements.

Material and methods. We used collagen-coated carriers (Cytodex3®) and a spinner flask bioreactor to develop environment suitable for human myoblast proliferation. In parallel, standard adherent in vitro culture conditions for myoblasts propagation (T-flask) were conducted. Cell cycle characterization, senescence, myogenic gene ex­pression and cell apoptosis were evaluated in order to find differences between two culture systems under study.

Results. The number of cells obtained in bioreactor per 106 of starting cells population was approximately ten times lower in comparison with T-flask culture system. The microcarriers cultured adult myoblasts in compari­son with the regular T-flask culture showed faster and more advanced replicative aging and lower proliferative potential. Moreover, the percentage of the cells that entailed an irreversible cell arrest (G0 phase) was also significantly (p < 0.0001) increased.

Conclusions. Our results suggest that population of primary human myoblasts obtained from adult individuals and propagated on dextran microcarriers did not meet the requirements of the regenerative medicine regarding quantity and quality of the cells obtained. Nonetheless, further optimization of the cell scaling up process including both microcarriers and/or bioreactor program is still an important option.

Abstract

Introduction. Primary cells in vitro culture scale-up is a crucial issue in cell-based tissue and organ regeneration therapy. Reducing costs and space occupied by the cells cultured in vitro has been an important target. Cells cultured in vitro with the use of bioreactor with dextran microcarriers (Cytodex®) have potentially a chance to meet many of the cell therapy requirements.

Material and methods. We used collagen-coated carriers (Cytodex3®) and a spinner flask bioreactor to develop environment suitable for human myoblast proliferation. In parallel, standard adherent in vitro culture conditions for myoblasts propagation (T-flask) were conducted. Cell cycle characterization, senescence, myogenic gene ex­pression and cell apoptosis were evaluated in order to find differences between two culture systems under study.

Results. The number of cells obtained in bioreactor per 106 of starting cells population was approximately ten times lower in comparison with T-flask culture system. The microcarriers cultured adult myoblasts in compari­son with the regular T-flask culture showed faster and more advanced replicative aging and lower proliferative potential. Moreover, the percentage of the cells that entailed an irreversible cell arrest (G0 phase) was also significantly (p < 0.0001) increased.

Conclusions. Our results suggest that population of primary human myoblasts obtained from adult individuals and propagated on dextran microcarriers did not meet the requirements of the regenerative medicine regarding quantity and quality of the cells obtained. Nonetheless, further optimization of the cell scaling up process including both microcarriers and/or bioreactor program is still an important option.

Get Citation

Keywords

human myoblasts; bioreactor; T-flask culture; stem cell precursors; cell senescence; telomeres; myogenic gene expression; cell cycle

About this article
Title

In vitro culture of primary human myoblasts by using the dextran microcarriers Cytodex3®

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 54, No 2 (2016)

Article type

Original paper

Pages

81-90

Published online

2016-06-07

DOI

10.5603/FHC.a2016.0010

Pubmed

27270505

Bibliographic record

Folia Histochem Cytobiol 2016;54(2):81-90.

Keywords

human myoblasts
bioreactor
T-flask culture
stem cell precursors
cell senescence
telomeres
myogenic gene expression
cell cycle

Authors

Natalia Rozwadowska
Agnieszka Malcher
Ewa Baumann
Tomasz Jan Kolanowski
Marek Rucinski
Tomasz Mietkiewski
Katarzyna Fiedorowicz
Maciej Kurpisz

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