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In vitro Myogenic Differentiation of Human Bone Marrow–Derived Mesenchymal Stem Cells as a Potential Treatment for Urethral Sphincter Muscle Repair
Author(s) -
Drost Adriana C.,
Weng Sibylle,
Feil Gerhard,
Schäfer Jochen,
Baumann Simon,
Kanz Lothar,
Sievert KarlDietrich,
Stenzl Arnulf,
Möhle Robert
Publication year - 2009
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1111/j.1749-6632.2009.04610.x
Subject(s) - mesenchymal stem cell , cd34 , cd31 , cd90 , cd146 , myosin , biology , antigen , stem cell , progenitor cell , microbiology and biotechnology , bone marrow , andrology , immunology , medicine , immunohistochemistry
To explore a new treatment strategy for urinary incontinence, human bone marrow mesenchymal stem cells (MSC) of the first in vitro passage were exposed to 5‐azacytidine (AZA) to induce myogenic differentiation, and cultured for a total of six passages. Expression of stem cell surface antigens and intracellular α‐actin was examined by flow cytometry at the end of each passage and compared to that of native MSC (not exposed to AZA) cultured in parallel. To analyze differentiation into striated muscle, expression of the transcription factor MyoD1 and myosin heavy chain (MyHC) was examined by RT‐PCR. Both native and AZA‐exposed MSC of all passages were negative for the progenitor/endothelial antigen CD34, leukocytic CD45, and endothelial/monocytic CD31. In contrast, the MSC markers CD73, CD90, CD105, and intracellular actin were detected in both groups of MSC throughout the culture period. After an initial increase, the expression level of MSC antigens decreased over time particularly in AZA‐exposed MSC. Expression of smooth muscle α‐actin also declined, but was greater in AZA‐exposed MSC throughout the culture period. Varying percentages of MSC cultures expressed MyoD1 and MyHC mRNA. In late passages, AZA‐exposed MSC tended to be more frequently positive than native MSC. In pilot experiments, transplantation of MSC into the bladder neck tissue of athymic rats was feasible; long‐term analyses are pending. We conclude that independent of AZA exposure, MSC express smooth and striated muscle antigens. Treatment with AZA slightly increases myogenic differentiation, but may not be necessary in future studies of MSC as a treatment modality for urinary incontinence.