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Differentiation of human umbilical cord matrix mesenchymal stem cells into neural-like progenitor cells and maturation into an oligodendroglial-like lineage.

Leite C, Silva NT, Mendes S, Ribeiro A, de Faria JP, Lourenço T, dos Santos F, Andrade PZ, Cardoso CM, Vieira M, Paiva A, da Silva CL, Cabral JM, Relvas JB, Grãos M - PLoS ONE (2014)

Bottom Line: In the present work MSCs were isolated from the umbilical cord matrix (Wharton's jelly) of human umbilical cord samples.The cells were additionally subjected to an oligodendroglial-oriented step-wise differentiation protocol in order to test their neural- and oligodendroglial-like differentiation capacity.The results confirmed the neural-like plasticity of MSCs, and suggested that the cells presented an oligodendroglial-like phenotype throughout the differentiation protocol, in several aspects sharing characteristics common to those of bona-fide oligodendrocyte precursor cells and differentiated oligodendrocytes.

View Article: PubMed Central - PubMed

Affiliation: Biocant - Technology Transfer Association, Biocant Park, Cantanhede, Portugal.

ABSTRACT
Mesenchymal stem cells (MSCs) are viewed as safe, readily available and promising adult stem cells, which are currently used in several clinical trials. Additionally, their soluble-factor secretion and multi-lineage differentiation capacities place MSCs in the forefront of stem cell types with expected near-future clinical applications. In the present work MSCs were isolated from the umbilical cord matrix (Wharton's jelly) of human umbilical cord samples. The cells were thoroughly characterized and confirmed as bona-fide MSCs, presenting in vitro low generation time, high proliferative and colony-forming unit-fibroblast (CFU-F) capacity, typical MSC immunophenotype and osteogenic, chondrogenic and adipogenic differentiation capacity. The cells were additionally subjected to an oligodendroglial-oriented step-wise differentiation protocol in order to test their neural- and oligodendroglial-like differentiation capacity. The results confirmed the neural-like plasticity of MSCs, and suggested that the cells presented an oligodendroglial-like phenotype throughout the differentiation protocol, in several aspects sharing characteristics common to those of bona-fide oligodendrocyte precursor cells and differentiated oligodendrocytes.

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Co-localization of MBP and F-actin in oligodendrocyte-like cells.Fluorescence microscopy images of OL-like cells (A) stained with fluorescent phalloidin (in green) to assess the presence of polymerized actin (F-actin) and an antibody against MBP (in red) and both channels merged, as indicated. Cells were derived from OPC-like cells cultured on FN-coated wells. Cells showing structured MBP and with partial co-localization with F-actin (arrow heads) or with diffuse MBP and no co-localization with F-actin (arrows) were visible after OL-like differentiation in the presence of T3, F3, T3+F3 or 7 days T3 followed by 3 days T3+F3 (T3+3d F3), as indicated. The bottom panel is a magnification of the T3 condition. Images are representative of at least 3 independent experiments and scale bar corresponds to 50 µm. The percentage of cells expressing structured MBP was calculated (B) under two categories: cells expressing structured cytoplasmic (left) or peripheral (right) MBP. Values were expressed as percentage of total cells and bars represent mean ± SEM percentage of cells present in at least 14 fields, belonging to 3 independent experiments. No statistically significant differences were found between the distinct differentiation conditions (Kruskal-Wallis one-way ANOVA followed by Dunn's multiple comparison test).
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pone-0111059-g008: Co-localization of MBP and F-actin in oligodendrocyte-like cells.Fluorescence microscopy images of OL-like cells (A) stained with fluorescent phalloidin (in green) to assess the presence of polymerized actin (F-actin) and an antibody against MBP (in red) and both channels merged, as indicated. Cells were derived from OPC-like cells cultured on FN-coated wells. Cells showing structured MBP and with partial co-localization with F-actin (arrow heads) or with diffuse MBP and no co-localization with F-actin (arrows) were visible after OL-like differentiation in the presence of T3, F3, T3+F3 or 7 days T3 followed by 3 days T3+F3 (T3+3d F3), as indicated. The bottom panel is a magnification of the T3 condition. Images are representative of at least 3 independent experiments and scale bar corresponds to 50 µm. The percentage of cells expressing structured MBP was calculated (B) under two categories: cells expressing structured cytoplasmic (left) or peripheral (right) MBP. Values were expressed as percentage of total cells and bars represent mean ± SEM percentage of cells present in at least 14 fields, belonging to 3 independent experiments. No statistically significant differences were found between the distinct differentiation conditions (Kruskal-Wallis one-way ANOVA followed by Dunn's multiple comparison test).

Mentions: Although we could not observe an increase of MBP expression in OL-like cells when compared with undifferentiated MSCs, it could be observed that there were distinct patterns of expression of MBP in the differentiated cells. One of the patterns was common to all stages of differentiation, including undifferentiated MSCs (Figure 6T), which was a strong staining in the nuclear area and more diffuse in cytosolic areas surrounding the nucleus (e.g.: Figure 8A, MBP panel, arrows). The other patterns suggested a structured distribution of MBP, resembling cytosolic filaments organized in a parallel manner, or at the periphery of the cells (e.g.: Figure 8A, MBP panel, arrow heads) and were present only in OL-like cells.


Differentiation of human umbilical cord matrix mesenchymal stem cells into neural-like progenitor cells and maturation into an oligodendroglial-like lineage.

Leite C, Silva NT, Mendes S, Ribeiro A, de Faria JP, Lourenço T, dos Santos F, Andrade PZ, Cardoso CM, Vieira M, Paiva A, da Silva CL, Cabral JM, Relvas JB, Grãos M - PLoS ONE (2014)

Co-localization of MBP and F-actin in oligodendrocyte-like cells.Fluorescence microscopy images of OL-like cells (A) stained with fluorescent phalloidin (in green) to assess the presence of polymerized actin (F-actin) and an antibody against MBP (in red) and both channels merged, as indicated. Cells were derived from OPC-like cells cultured on FN-coated wells. Cells showing structured MBP and with partial co-localization with F-actin (arrow heads) or with diffuse MBP and no co-localization with F-actin (arrows) were visible after OL-like differentiation in the presence of T3, F3, T3+F3 or 7 days T3 followed by 3 days T3+F3 (T3+3d F3), as indicated. The bottom panel is a magnification of the T3 condition. Images are representative of at least 3 independent experiments and scale bar corresponds to 50 µm. The percentage of cells expressing structured MBP was calculated (B) under two categories: cells expressing structured cytoplasmic (left) or peripheral (right) MBP. Values were expressed as percentage of total cells and bars represent mean ± SEM percentage of cells present in at least 14 fields, belonging to 3 independent experiments. No statistically significant differences were found between the distinct differentiation conditions (Kruskal-Wallis one-way ANOVA followed by Dunn's multiple comparison test).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4214693&req=5

pone-0111059-g008: Co-localization of MBP and F-actin in oligodendrocyte-like cells.Fluorescence microscopy images of OL-like cells (A) stained with fluorescent phalloidin (in green) to assess the presence of polymerized actin (F-actin) and an antibody against MBP (in red) and both channels merged, as indicated. Cells were derived from OPC-like cells cultured on FN-coated wells. Cells showing structured MBP and with partial co-localization with F-actin (arrow heads) or with diffuse MBP and no co-localization with F-actin (arrows) were visible after OL-like differentiation in the presence of T3, F3, T3+F3 or 7 days T3 followed by 3 days T3+F3 (T3+3d F3), as indicated. The bottom panel is a magnification of the T3 condition. Images are representative of at least 3 independent experiments and scale bar corresponds to 50 µm. The percentage of cells expressing structured MBP was calculated (B) under two categories: cells expressing structured cytoplasmic (left) or peripheral (right) MBP. Values were expressed as percentage of total cells and bars represent mean ± SEM percentage of cells present in at least 14 fields, belonging to 3 independent experiments. No statistically significant differences were found between the distinct differentiation conditions (Kruskal-Wallis one-way ANOVA followed by Dunn's multiple comparison test).
Mentions: Although we could not observe an increase of MBP expression in OL-like cells when compared with undifferentiated MSCs, it could be observed that there were distinct patterns of expression of MBP in the differentiated cells. One of the patterns was common to all stages of differentiation, including undifferentiated MSCs (Figure 6T), which was a strong staining in the nuclear area and more diffuse in cytosolic areas surrounding the nucleus (e.g.: Figure 8A, MBP panel, arrows). The other patterns suggested a structured distribution of MBP, resembling cytosolic filaments organized in a parallel manner, or at the periphery of the cells (e.g.: Figure 8A, MBP panel, arrow heads) and were present only in OL-like cells.

Bottom Line: In the present work MSCs were isolated from the umbilical cord matrix (Wharton's jelly) of human umbilical cord samples.The cells were additionally subjected to an oligodendroglial-oriented step-wise differentiation protocol in order to test their neural- and oligodendroglial-like differentiation capacity.The results confirmed the neural-like plasticity of MSCs, and suggested that the cells presented an oligodendroglial-like phenotype throughout the differentiation protocol, in several aspects sharing characteristics common to those of bona-fide oligodendrocyte precursor cells and differentiated oligodendrocytes.

View Article: PubMed Central - PubMed

Affiliation: Biocant - Technology Transfer Association, Biocant Park, Cantanhede, Portugal.

ABSTRACT
Mesenchymal stem cells (MSCs) are viewed as safe, readily available and promising adult stem cells, which are currently used in several clinical trials. Additionally, their soluble-factor secretion and multi-lineage differentiation capacities place MSCs in the forefront of stem cell types with expected near-future clinical applications. In the present work MSCs were isolated from the umbilical cord matrix (Wharton's jelly) of human umbilical cord samples. The cells were thoroughly characterized and confirmed as bona-fide MSCs, presenting in vitro low generation time, high proliferative and colony-forming unit-fibroblast (CFU-F) capacity, typical MSC immunophenotype and osteogenic, chondrogenic and adipogenic differentiation capacity. The cells were additionally subjected to an oligodendroglial-oriented step-wise differentiation protocol in order to test their neural- and oligodendroglial-like differentiation capacity. The results confirmed the neural-like plasticity of MSCs, and suggested that the cells presented an oligodendroglial-like phenotype throughout the differentiation protocol, in several aspects sharing characteristics common to those of bona-fide oligodendrocyte precursor cells and differentiated oligodendrocytes.

Show MeSH
Related in: MedlinePlus