Maintenance of differentiation potential of human bone marrow mesenchymal stem cells immortalized by human telomerase reverse transcriptase gene despite of extensive proliferation

Abdallah, Basem M.; Haack‐Sørensen, Mandana; Burns, Jorge S.; Elsnab, Birgitte; Jakob, Franz; Hokland, Peter; Kassem, Moustapha

doi:10.1016/j.bbrc.2004.11.059

Cited by 234 publications

(215 citation statements)

References 44 publications

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“…1C). Increased expression of osteoblast-associated genes and the observed osteoblast phenotype were in accordance with previous reports describing hMSC differentiation into osteoblasts (1)(2)(3).…”

Section: Identification Of Differentially Expressed Mirnas During Ostsupporting

confidence: 92%

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MicroRNA-138 regulates osteogenic differentiation of human stromal (mesenchymal) stem cells in vivo

Eskildsen

Taipaleenmäki

Stenvang

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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443

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Elucidating the molecular mechanisms that regulate human stromal (mesenchymal) stem cell (hMSC) differentiation into osteogenic lineage is important for the development of anabolic therapies for treatment of osteoporosis. MicroRNAs (miRNAs) are short, noncoding RNAs that act as key regulators of diverse biological processes by mediating translational repression or mRNA degradation of their target genes. Here, we show that miRNA-138 (miR-138) modulates osteogenic differentiation of hMSCs. miRNA array profiling and further validation by quantitative RT-PCR (qRT-PCR) revealed that miR-138 was down-regulated during osteoblast differentiation of hMSCs. Overexpression of miR-138 inhibited osteoblast differentiation of hMSCs in vitro, whereas inhibition of miR-138 function by antimiR-138 promoted expression of osteoblast-specific genes, alkaline phosphatase (ALP) activity, and matrix mineralization. Furthermore, overexpression of miR-138 reduced ectopic bone formation in vivo by 85%, and conversely, in vivo bone formation was enhanced by 60% when miR-138 was antagonized. Target prediction analysis and experimental validation by luciferase 3′ UTR reporter assay confirmed focal adhesion kinase, a kinase playing a central role in promoting osteoblast differentiation, as a bona fide target of miR-138. We show that miR-138 attenuates bone formation in vivo, at least in part by inhibiting the focal adhesion kinase signaling pathway. Our findings suggest that pharmacological inhibition of miR-138 by antimiR-138 could represent a therapeutic strategy for enhancing bone formation in vivo.regulatory RNA | bone biology | osteoblastic differentiation

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Section: Identification Of Differentially Expressed Mirnas During Ostsupporting

confidence: 92%

“…hMSCs can differentiate along several lineages, including the osteogenic lineage, in response to stimulation by multiple environmental factors (1)(2)(3)(4), and they involve complex pathways regulated at both transcriptional and posttranscriptional levels. However, the regulation of these cellular pathways is not fully understood.…”

mentioning

confidence: 99%

MicroRNA-138 regulates osteogenic differentiation of human stromal (mesenchymal) stem cells in vivo

Eskildsen

Taipaleenmäki

Stenvang

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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443

354

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“…Establishment and culture conditions for telemorase immortalized human mesenchymal stem cells (hMSC-TERT) have been described in previous reports (Abdallah et al, 2005;Simonsen et al, 2002). Bone marrow (BM) aspirates were obtained from the posterior iliac crest of normal adult volunteers (19-35 years old) following informed consent, according to procedures approved by the ethics committee of the Royal Adelaide Hospital, South Australia.…”

Section: Cell Lines and Cell Culture Conditionsmentioning

confidence: 99%

“…In order to avoid culture and donor variations of hMSC, we employed an immortalized human telomerase reverse transcriptase transduced MSC line (hMSC-TERT) that has been developed by our group and expresses typical characertistics of hMSC both in vitro and in vivo (Abdallah et al, 2005;Simonsen et al, 2002). hMSC-TERT cells were thus employed to immunize mice for antibody development, where human skin fibroblasts were used for negative selection to enhance antibody specificity by eliminating common markers with hMSC.…”

Section: Introductionmentioning

confidence: 99%

Development of Novel Monoclonal Antibodies that Define Differentiation Stages of Human Stromal (Mesenchymal) Stem Cells

Andersen

Kortesidis

Zannettino

et al. 2011

Molecules and Cells

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Human mesenchymal stem cells (hMSC) are currently being introduced for cell therapy, yet, antibodies specific for native and differentiated MSCs are required for their identification prior to clinical use. Herein, high quality antibodies against MSC surface proteins were developed by immunizing mice with hMSC, and by using a panel of subsequent screening methods. Flow cytometry analysis revealed that 83.5, 1.1, and 8.5% of primary cultures of hMSC were double positive for STRO-1 and either of DJ 3, 9, and 18, respectively. However, none of the three DJ antibodies allowed enrichment of clonogenic hMSC from BMMNCs as single reagents. Using mass-spectrometric analysis, we identified the antigen recognised by DJ3 as CD44, whereas DJ9 and DJ18 recognized HLA-DRB1 and Collagen VI, respectively. The identified proteins were highly expressed throughout in vitro osteogenic-and adipogenic differentiation. Interestingly, undifferentiated cells revealed a sole cytoplasmic distribution pattern of Collagen VI, which however changed to an extracellular matrix appearance upon osteogenic-and adipogenic differentiation. In relation to this, we found that STRO-1 +/-/Collagen VI -sorted hMSC contained fewer differentiated alkaline phosphatase + cells compared to STRO-1 +/-/Collagen VI + hMSC, suggesting that Collagen VI on the cell membrane exclusively defines differentiated MSCs. In conclusion, we have generated a panel of high quality antibodies to be used for characterization of MSCs, and in addition our results may suggest that the DJ18 generated antibody against Collagen VI can be used for negative selection of cultured undifferentiated MSCs.

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“…[15][16][17][18][19] The phenotypic changes in BM-MSCs in culture are differentiation, dedifferentiation, replicative senescence, and spontaneous transformation, and studies designed to overcome these changes have been pursued. [20][21][22][23][24] However, few studies have been undertaken on synoviumderived MSCs.…”

mentioning

confidence: 99%

Changes in chondrogenic phenotype and gene expression profiles associated with the in vitro expansion of human synovium‐derived cells

Han

Lee

Kim

et al. 2010

Journal Orthopaedic Research

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We undertook this study to characterize changes in the proliferative capacities, chondrogenic phenotypes, and gene expression profiles of human synovium-derived progenitor cells from osteoarthritic patients during in vitro expansion. Cells isolated from osteoarthritic synovia were cultured, and growth rates during serial passages were evaluated. Surface molecule expressions were determined by flow cytometry and cytogenetic analyses were performed. After chondrogenic differentiation in cell pellets, we evaluated type II collagen and glycosaminoglycan (GAG) synthesis. To assess whether the in vitro expansion of synovium-derived cells affects gene expression, we performed microarray analyses on cells at passage 0, 1, 2, 4, 6, and 8. Synovium-derived cells were rapidly expanded in vitro through passage 8 (about 130 days), and after passage 6, the proliferation rates decreased slightly with a wide range of individual variations. The expressions of CD166, CD49a, and CD106 decreased, whereas those of CD10, CD29, CD44, CD73, CD90, and CD105 showed no significant change. Karyotype analysis revealed no evidence of chromosome abnormalities. The staining of type II collagen and GAG in differentiated cell pellets showed rapid weakening. Genome-wide microarray analysis showed that synovium-derived cells from late passages over-expressed genes associated with cell cycle prolongation and cell aging, and less-expressed genes associated with cell growth stimulation. The in vitro expansion of synovium-derived cells was accompanied with decreased proliferative capacity and the chondrogenic phenotype, which might be modulated by change in gene expression patterns. ß

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Maintenance of differentiation potential of human bone marrow mesenchymal stem cells immortalized by human telomerase reverse transcriptase gene despite of extensive proliferation

Cited by 234 publications

References 44 publications

MicroRNA-138 regulates osteogenic differentiation of human stromal (mesenchymal) stem cells in vivo

MicroRNA-138 regulates osteogenic differentiation of human stromal (mesenchymal) stem cells in vivo

Development of Novel Monoclonal Antibodies that Define Differentiation Stages of Human Stromal (Mesenchymal) Stem Cells

Changes in chondrogenic phenotype and gene expression profiles associated with the in vitro expansion of human synovium‐derived cells

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