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Generation of mesenchymal stem cell lines from murine bone marrow
Dillip Kumar Bishi,
Published in
2012
PMID: 22836234
Volume: 350
   
Issue: 1
Pages: 55 - 68
Abstract
Mesenchymal stem cells (MSC), because of their multipotency and ease of purification and amplification, are an ideal stem cell source for cell therapies. Bone-marrow-derived stem cells (BMSC) can be used to develop MSC-like immortalized cell lines with large proliferation and differentiation potentialities. Their immortalized status prevents the maintenance of MSC function and characters; this can be negated by modifying the isolation and maintenance protocol. Adult murine BMSC were isolated and maintained in media without additional growth factors together with passage-dependent reseeding following trypsinization. Cells maintained over 25 passages were considered as putative cell lines and characterized. The phenotypic and genotypic characteristics and multilineage differentiation potential of the cells were assessed by morphological, phenotypic, and molecular assays at various passages. The putative BMSC cell lines showed the characteristics of MSC and were able to maintain these characteristics, even after immortalization. The phenotypic data demonstrated difference among two cell lines; this was further validated by the difference in their multilineage differentiation potential following specific induction. More importantly, no changes were observed in the genotypic level in comparison with control cells, even after more than 50 passages. Our protocol thus advances the isolation and maintenance of BMSC and the development of putative BMSC cell lines that maintain characteristics of MSC, including multilineage differentiation potential, after more than 40 passages. © 2012 Springer-Verlag.
About the journal
JournalCell and Tissue Research
ISSN0302766X
Open AccessNo
Concepts (44)
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    ADIPOGENESIS
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    Animal cell
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    Animal cell culture
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    Article
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    Biosynthesis
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    CARDIOMYOGENESIS
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    Cell assay
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    Cell differentiation
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    CELL IMMORTALIZATION
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    CELL ISOLATION
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    Cell lineage
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    Cell population
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    Clinical protocol
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    Controlled study
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    Fibroblast
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    FLUORESCENCE ACTIVATED CELL SORTING
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    Genotype
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    HEMATOPOIETIC STEM CELL
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    IMMUNOCYTOCHEMISTRY
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    Mesenchymal stem cell
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    Mouse
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    Nonhuman
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    OSTEOBLASTOGENESIS
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    Phenotype
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    Priority journal
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    Process development
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    Purification
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    Reproducibility
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    ADIPOGENESIS
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    Animals
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    Bone marrow cells
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    Cell culture techniques
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    Cell cycle
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    Cell line
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    Flow cytometry
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    Immunohistochemistry
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    KARYOTYPING
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    Mesenchymal stromal cells
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    Mice
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    MICROSCOPY, PHASE-CONTRAST
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    MYOCYTES, CARDIAC
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    OSTEOGENESIS
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    Reverse transcriptase polymerase chain reaction
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    MURINAE