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Role of Slug transcription factor in human mesenchymal stem cells.

Torreggiani E, Lisignoli G, Manferdini C, Lambertini E, Penolazzi L, Vecchiatini R, Gabusi E, Chieco P, Facchini A, Gambari R, Piva R - J. Cell. Mol. Med. (2012)

Bottom Line: In this study we found that Slug-silenced cells changed in morphology, decreased in their migration ability, increased Sox9 and Sox5 and decreased Sox6 and STAT1 expression.On the contrary, the effect of Slug depletion on Runx2 was influenced by cell type.As a whole, our findings have important potential implication on bone tissue engineering applications, reinforcing the concept that manipulation of specific TF expression levels may elucidate MSC biology and the molecular mechanisms, which promote osteogenic differentiation.

View Article: PubMed Central - PubMed

Affiliation: Dipartimento di Biochimica e Biologia Molecolare, Sezione di Biologia Molecolare, Università degli Studi di Ferrara, Ferrara, Italy.

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Osteogenic differentiation of hMSCs. (A) Differentiation potential of hMSCs from the three sources was assessed by evaluating the mineralization of the cell cultures in the presence of osteogenic induction medium, up to 21 days. Data were expressed as integrated optical density (O.D.) ± S.E. Statistical analysis was performed hTP-MSC versus hWJ-MSC*, hTP-MSC versus hIC-MSC^ and hIC-MSC versus hWJ-MSCo, as described in Results. (B) Slug, Runx2, Sox9, Sox5, Sox6 and STAT1 gene expression was valuated in hMSCs isolated from tibial plateau (TP) trabecular bone, iliac crest (IC) bone marrow and Wharton’s jelly (WJ) umbilical cord and cultured up to 21 days in osteogenic medium. mRNA level was revealed by quantitative RT-PCR analysis. RT-PCR results were calculated using the DDCt method. Data were expressed as fold difference value between the calibrator (Sox9 expression level at day 21 in hWJ-MSCs) and the other samples. Data were expressed as median ± S.E.M. It was not possible to apply statistical tests because the N was equal to four samples for each group analysed.
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fig06: Osteogenic differentiation of hMSCs. (A) Differentiation potential of hMSCs from the three sources was assessed by evaluating the mineralization of the cell cultures in the presence of osteogenic induction medium, up to 21 days. Data were expressed as integrated optical density (O.D.) ± S.E. Statistical analysis was performed hTP-MSC versus hWJ-MSC*, hTP-MSC versus hIC-MSC^ and hIC-MSC versus hWJ-MSCo, as described in Results. (B) Slug, Runx2, Sox9, Sox5, Sox6 and STAT1 gene expression was valuated in hMSCs isolated from tibial plateau (TP) trabecular bone, iliac crest (IC) bone marrow and Wharton’s jelly (WJ) umbilical cord and cultured up to 21 days in osteogenic medium. mRNA level was revealed by quantitative RT-PCR analysis. RT-PCR results were calculated using the DDCt method. Data were expressed as fold difference value between the calibrator (Sox9 expression level at day 21 in hWJ-MSCs) and the other samples. Data were expressed as median ± S.E.M. It was not possible to apply statistical tests because the N was equal to four samples for each group analysed.

Mentions: We then investigated the osteogenic potential of hMSCs from the three sources. At 21 days of differentiation, the cultures showed the presence of mineralized nodules following alizarin red staining analysis (Fig. 6A). These data support the hypothesis that MSCs can be successfully differentiated towards osteogenic lineage when appropriately stimulated in vitro. Interestingly, mineralization occurred at day 7 only for hWJ-MSC whereas for hTP- and hIC-MSC started from day 14. In addition, we noticed that, both at day 14 and 21, mineralization was significantly increased in hTP-MSC (P = 0.003 and 0.006, respectively) and hIC-MSC (P = 0.00001 and 0.006, respectively) compared to hWJ-MSC. Comparing hTP- and hIC-MSC we observed that, whereas at day 14 mineralization was significantly higher in hIC-MSC than in hTP-MSC (P = 0.008), at day 21 it reached approximately the same values in both cell sources.


Role of Slug transcription factor in human mesenchymal stem cells.

Torreggiani E, Lisignoli G, Manferdini C, Lambertini E, Penolazzi L, Vecchiatini R, Gabusi E, Chieco P, Facchini A, Gambari R, Piva R - J. Cell. Mol. Med. (2012)

Osteogenic differentiation of hMSCs. (A) Differentiation potential of hMSCs from the three sources was assessed by evaluating the mineralization of the cell cultures in the presence of osteogenic induction medium, up to 21 days. Data were expressed as integrated optical density (O.D.) ± S.E. Statistical analysis was performed hTP-MSC versus hWJ-MSC*, hTP-MSC versus hIC-MSC^ and hIC-MSC versus hWJ-MSCo, as described in Results. (B) Slug, Runx2, Sox9, Sox5, Sox6 and STAT1 gene expression was valuated in hMSCs isolated from tibial plateau (TP) trabecular bone, iliac crest (IC) bone marrow and Wharton’s jelly (WJ) umbilical cord and cultured up to 21 days in osteogenic medium. mRNA level was revealed by quantitative RT-PCR analysis. RT-PCR results were calculated using the DDCt method. Data were expressed as fold difference value between the calibrator (Sox9 expression level at day 21 in hWJ-MSCs) and the other samples. Data were expressed as median ± S.E.M. It was not possible to apply statistical tests because the N was equal to four samples for each group analysed.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3822845&req=5

fig06: Osteogenic differentiation of hMSCs. (A) Differentiation potential of hMSCs from the three sources was assessed by evaluating the mineralization of the cell cultures in the presence of osteogenic induction medium, up to 21 days. Data were expressed as integrated optical density (O.D.) ± S.E. Statistical analysis was performed hTP-MSC versus hWJ-MSC*, hTP-MSC versus hIC-MSC^ and hIC-MSC versus hWJ-MSCo, as described in Results. (B) Slug, Runx2, Sox9, Sox5, Sox6 and STAT1 gene expression was valuated in hMSCs isolated from tibial plateau (TP) trabecular bone, iliac crest (IC) bone marrow and Wharton’s jelly (WJ) umbilical cord and cultured up to 21 days in osteogenic medium. mRNA level was revealed by quantitative RT-PCR analysis. RT-PCR results were calculated using the DDCt method. Data were expressed as fold difference value between the calibrator (Sox9 expression level at day 21 in hWJ-MSCs) and the other samples. Data were expressed as median ± S.E.M. It was not possible to apply statistical tests because the N was equal to four samples for each group analysed.
Mentions: We then investigated the osteogenic potential of hMSCs from the three sources. At 21 days of differentiation, the cultures showed the presence of mineralized nodules following alizarin red staining analysis (Fig. 6A). These data support the hypothesis that MSCs can be successfully differentiated towards osteogenic lineage when appropriately stimulated in vitro. Interestingly, mineralization occurred at day 7 only for hWJ-MSC whereas for hTP- and hIC-MSC started from day 14. In addition, we noticed that, both at day 14 and 21, mineralization was significantly increased in hTP-MSC (P = 0.003 and 0.006, respectively) and hIC-MSC (P = 0.00001 and 0.006, respectively) compared to hWJ-MSC. Comparing hTP- and hIC-MSC we observed that, whereas at day 14 mineralization was significantly higher in hIC-MSC than in hTP-MSC (P = 0.008), at day 21 it reached approximately the same values in both cell sources.

Bottom Line: In this study we found that Slug-silenced cells changed in morphology, decreased in their migration ability, increased Sox9 and Sox5 and decreased Sox6 and STAT1 expression.On the contrary, the effect of Slug depletion on Runx2 was influenced by cell type.As a whole, our findings have important potential implication on bone tissue engineering applications, reinforcing the concept that manipulation of specific TF expression levels may elucidate MSC biology and the molecular mechanisms, which promote osteogenic differentiation.

View Article: PubMed Central - PubMed

Affiliation: Dipartimento di Biochimica e Biologia Molecolare, Sezione di Biologia Molecolare, Università degli Studi di Ferrara, Ferrara, Italy.

Show MeSH
Related in: MedlinePlus