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Characterisation of nuclear architectural alterations during in vitro differentiation of human stem cells of myogenic origin.

Rozwadowska N, Kolanowski T, Wiland E, Siatkowski M, Pawlak P, Malcher A, Mietkiewski T, Olszewska M, Kurpisz M - PLoS ONE (2013)

Bottom Line: Additionally, almost all of the chromosomal centromeres in the differentiated cells preferentially localised near the nuclear periphery when compared to the undifferentiated cells.The exceptions were chromosomes 7 and 11, in which we were unable to confirm the centromere repositioning.Based on these data we can conclude that the myogenic differentiation with global gene expression changes is accompanied by the spatial repositioning of chromosomes and chromatin remodelling, which are important processes that regulate cell differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Reproductive Biology and Stem Cells, Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland.

ABSTRACT
Cell differentiation is based on a synchronised orchestra of complex pathways of intrinsic and extrinsic signals that manifest in the induced expression of specific transcription factors and pivotal genes within the nucleus. One cannot ignore the epigenetic status of differentiating cells, comprising not only histones and DNA modifications but also the spatial and temporal intranuclear chromatin organisation, which is an important regulator of nuclear processes. In the present study, we investigated the nuclear architecture of human primary myoblasts and myocytes in an in vitro culture, with reference to global changes in genomic expression. Repositioning of the chromosomal centromeres, along with alterations in the nuclear shape and volume, was observed as a consequence of myotube formation. Moreover, the microarray data showed that during in vitro myogenesis cells tend to silence rather than induce gene expression. The creation of a chromosome map marked with gene expression changes that were at least 2-fold confirmed the observation. Additionally, almost all of the chromosomal centromeres in the differentiated cells preferentially localised near the nuclear periphery when compared to the undifferentiated cells. The exceptions were chromosomes 7 and 11, in which we were unable to confirm the centromere repositioning. In our opinion, this is the first reported observation of the movement of chromosomal centromeres along differentiating myogenic cells. Based on these data we can conclude that the myogenic differentiation with global gene expression changes is accompanied by the spatial repositioning of chromosomes and chromatin remodelling, which are important processes that regulate cell differentiation.

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Human myoblasts in in vitro culture.A – flow cytometry analysis of CD56 expression on human myoblasts. Mean percent of cells positive for the CD56 marker: 85.0+/−5.0%. Panel B - in vitro culture of myoblasts and myocytes (Mb24 hrs – proliferating myoblasts; Mc7d- fused myocytes (myotubes). magnification-100x).
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pone-0073231-g002: Human myoblasts in in vitro culture.A – flow cytometry analysis of CD56 expression on human myoblasts. Mean percent of cells positive for the CD56 marker: 85.0+/−5.0%. Panel B - in vitro culture of myoblasts and myocytes (Mb24 hrs – proliferating myoblasts; Mc7d- fused myocytes (myotubes). magnification-100x).

Mentions: Isolated myoblasts were tested for their differentiation potential and characterised as CD56+positive when expression levels between 80%–90% were observed for the cell populations under study (Figure 2A). The morphology of cells that had differentiated into myocytes changed rapidly, from fibroblast-like mononuclear cells to strongly elongated fused multinucleated myotubes (Figure 2B). The calculated fusion index showed efficient in vitro myocytes formation (Fi = 4.5±0.25). (Figure S1).


Characterisation of nuclear architectural alterations during in vitro differentiation of human stem cells of myogenic origin.

Rozwadowska N, Kolanowski T, Wiland E, Siatkowski M, Pawlak P, Malcher A, Mietkiewski T, Olszewska M, Kurpisz M - PLoS ONE (2013)

Human myoblasts in in vitro culture.A – flow cytometry analysis of CD56 expression on human myoblasts. Mean percent of cells positive for the CD56 marker: 85.0+/−5.0%. Panel B - in vitro culture of myoblasts and myocytes (Mb24 hrs – proliferating myoblasts; Mc7d- fused myocytes (myotubes). magnification-100x).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3760906&req=5

pone-0073231-g002: Human myoblasts in in vitro culture.A – flow cytometry analysis of CD56 expression on human myoblasts. Mean percent of cells positive for the CD56 marker: 85.0+/−5.0%. Panel B - in vitro culture of myoblasts and myocytes (Mb24 hrs – proliferating myoblasts; Mc7d- fused myocytes (myotubes). magnification-100x).
Mentions: Isolated myoblasts were tested for their differentiation potential and characterised as CD56+positive when expression levels between 80%–90% were observed for the cell populations under study (Figure 2A). The morphology of cells that had differentiated into myocytes changed rapidly, from fibroblast-like mononuclear cells to strongly elongated fused multinucleated myotubes (Figure 2B). The calculated fusion index showed efficient in vitro myocytes formation (Fi = 4.5±0.25). (Figure S1).

Bottom Line: Additionally, almost all of the chromosomal centromeres in the differentiated cells preferentially localised near the nuclear periphery when compared to the undifferentiated cells.The exceptions were chromosomes 7 and 11, in which we were unable to confirm the centromere repositioning.Based on these data we can conclude that the myogenic differentiation with global gene expression changes is accompanied by the spatial repositioning of chromosomes and chromatin remodelling, which are important processes that regulate cell differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Reproductive Biology and Stem Cells, Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland.

ABSTRACT
Cell differentiation is based on a synchronised orchestra of complex pathways of intrinsic and extrinsic signals that manifest in the induced expression of specific transcription factors and pivotal genes within the nucleus. One cannot ignore the epigenetic status of differentiating cells, comprising not only histones and DNA modifications but also the spatial and temporal intranuclear chromatin organisation, which is an important regulator of nuclear processes. In the present study, we investigated the nuclear architecture of human primary myoblasts and myocytes in an in vitro culture, with reference to global changes in genomic expression. Repositioning of the chromosomal centromeres, along with alterations in the nuclear shape and volume, was observed as a consequence of myotube formation. Moreover, the microarray data showed that during in vitro myogenesis cells tend to silence rather than induce gene expression. The creation of a chromosome map marked with gene expression changes that were at least 2-fold confirmed the observation. Additionally, almost all of the chromosomal centromeres in the differentiated cells preferentially localised near the nuclear periphery when compared to the undifferentiated cells. The exceptions were chromosomes 7 and 11, in which we were unable to confirm the centromere repositioning. In our opinion, this is the first reported observation of the movement of chromosomal centromeres along differentiating myogenic cells. Based on these data we can conclude that the myogenic differentiation with global gene expression changes is accompanied by the spatial repositioning of chromosomes and chromatin remodelling, which are important processes that regulate cell differentiation.

Show MeSH
Related in: MedlinePlus