Myogenic differentiation of mesenchymal stem cells co-cultured with primary myoblasts.
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Myogenic differentiation of mesenchymal stem cells co-cultured with primary myoblasts. / Beier, Justus P; Bitto, Franz F; Lange, Claudia; Klumpp, Dorothee; Arkudas, Andreas; Bleiziffer, Oliver; Boos, Anja M; Horch, Raymund E; Kneser, Ulrich.
In: CELL BIOL INT, Vol. 35, No. 4, 4, 2011, p. 397-406.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Myogenic differentiation of mesenchymal stem cells co-cultured with primary myoblasts.
AU - Beier, Justus P
AU - Bitto, Franz F
AU - Lange, Claudia
AU - Klumpp, Dorothee
AU - Arkudas, Andreas
AU - Bleiziffer, Oliver
AU - Boos, Anja M
AU - Horch, Raymund E
AU - Kneser, Ulrich
PY - 2011
Y1 - 2011
N2 - TE (tissue engineering) of skeletal muscle is a promising method to reconstruct loss of muscle tissue. This study evaluates MSCs (mesenchymal stem cells) as new cell source for this application. As a new approach to differentiate the MSCs towards the myogenic lineage, co-cultivation with primary myoblasts has been developed and the myogenic potential of GFP (green fluorescent protein)-transduced rat MSC co-cultured with primary rat myoblasts was assessed by ICC (immunocytochemistry). Myogenic potential of MSC was analysed by ICC, FACS and qPCR (quantitative PCR). MSC-myoblast fusion phenomena leading to hybrid myotubes were evaluated using a novel method to evaluate myotube fusion ratios based on phase contrast and fluorescence microscopy. Furthermore, MSC constitutively expressed the myogenic markers MEF2 (myogenic enhancer factor 2) and ?-sarcomeric actin, and MEF2 expression was up-regulated upon co-cultivation with primary myoblasts and the addition of myogenic medium supplements. Significantly higher numbers of MSC nuclei were involved in myotube formations when bFGF (basic fibroblast growth factor) and dexamethasone were added to co-cultures. In summary, we have determined optimal co-culture conditions for MSC myogenic differentiation up to myotube formations as a promising step towards applicability of MSC as a cell source for skeletal muscle TE as well as other muscle cell-based therapies.
AB - TE (tissue engineering) of skeletal muscle is a promising method to reconstruct loss of muscle tissue. This study evaluates MSCs (mesenchymal stem cells) as new cell source for this application. As a new approach to differentiate the MSCs towards the myogenic lineage, co-cultivation with primary myoblasts has been developed and the myogenic potential of GFP (green fluorescent protein)-transduced rat MSC co-cultured with primary rat myoblasts was assessed by ICC (immunocytochemistry). Myogenic potential of MSC was analysed by ICC, FACS and qPCR (quantitative PCR). MSC-myoblast fusion phenomena leading to hybrid myotubes were evaluated using a novel method to evaluate myotube fusion ratios based on phase contrast and fluorescence microscopy. Furthermore, MSC constitutively expressed the myogenic markers MEF2 (myogenic enhancer factor 2) and ?-sarcomeric actin, and MEF2 expression was up-regulated upon co-cultivation with primary myoblasts and the addition of myogenic medium supplements. Significantly higher numbers of MSC nuclei were involved in myotube formations when bFGF (basic fibroblast growth factor) and dexamethasone were added to co-cultures. In summary, we have determined optimal co-culture conditions for MSC myogenic differentiation up to myotube formations as a promising step towards applicability of MSC as a cell source for skeletal muscle TE as well as other muscle cell-based therapies.
KW - Animals
KW - Male
KW - Cells, Cultured
KW - Rats
KW - Cell Line
KW - Tissue Engineering/methods
KW - Cell Differentiation
KW - Coculture Techniques/methods
KW - Mesenchymal Stem Cells/cytology
KW - Myoblasts/cytology
KW - Rats, Inbred Lew
KW - Animals
KW - Male
KW - Cells, Cultured
KW - Rats
KW - Cell Line
KW - Tissue Engineering/methods
KW - Cell Differentiation
KW - Coculture Techniques/methods
KW - Mesenchymal Stem Cells/cytology
KW - Myoblasts/cytology
KW - Rats, Inbred Lew
M3 - SCORING: Journal article
VL - 35
SP - 397
EP - 406
JO - CELL BIOL INT
JF - CELL BIOL INT
SN - 1065-6995
IS - 4
M1 - 4
ER -