In vivo and in vitro differentiation of myocytes from human bone marrow-derived multipotent progenitor cells.
OBJECTIVE
Recent studies have shown that bone marrow (BM) contains cells capable of differentiating into myocytes in vivo. However, addition of demethylation drugs has been necessary to induce myocyte differentiation from BM cells in vitro, and precise mechanisms of BM cells' conversion to myocytes and the origin of those cells have not been established. We investigated the expression of myogenic markers during differentiation and maturation of myocytes from BM-derived multipotent adult progenitor cells (MAPC) under physiological culture condition.


METHODS
Frozen BM samples from 21 healthy donors were used as a source of MAPC. To induce myocyte differentiation MAPC was cultured in the presence of 5% FCS, VEGF, bFGF, and IGF-1, and the expressions of myocyte markers were examined at various time points. We also investigated engraftment and differentiation of MAPC-derived myocytes in vivo.


RESULTS
Frozen BM-derived MAPC, cultured under the physiological myogenic condition, demonstrated spatial expression patterns of several myocyte markers similar to that of authentic myocyte differentiation. When injected into murine muscles, MAPC treated with the myogenic condition engrafted and differentiated into myocyte marker-positive cells and myotubes in vivo.


CONCLUSIONS
For the first time, we were able to induce myocyte formation from BM cells under the physiological condition in vitro and demonstrated that treating cells with this condition prior to intramuscular injection increased efficiency of engraftment and differentiation in vivo.
Muguruma
Yukari
Y
Division of Hematopoiesis, Research Center for Regenerative Medicine, Tokai University School of Medicine, Boseidai, Isehara, Kanagawa 259-1193, Japan.
Reyes
Morayma
M
Nakamura
Yoshihiko
Y
Sato
Tadayuki
T
Matsuzawa
Hideyuki
H
Miyatake
Hiroko
H
Akatsuka
Akira
A
Itoh
Johbu
J
Yahata
Takashi
T
Ando
Kiyoshi
K
Kato
Shunichi
S
Hotta
Tomomitsu
T
D000293:Adolescent; D000328:Adult; D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D001854:Bone Marrow Cells; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D019070:Cell Lineage; D017690:Cell Transplantation; D002648:Child; D006085:Graft Survival; D006801:Humans; D051379:Mice; D008875:Middle Aged; D039902:Multipotent Stem Cells / Q000166:cytology*; D032342:Muscle Cells / Q000166:cytology* / Q000637:transplantation*; D018485:Muscle Fibers, Skeletal / Q000166:cytology; D014183:Transplantation, Heterologous
D015703:Antigens, CD