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Retinoic Acid as the Stimulating Factor for Differentiation of Wharton's Jelly-Mesenchymal Stem Cells into Hepatocyte-like Cells.

Mortezaee K, Minaii B, Sabbaghziarani F, Ragerdi Kashani I, Hassanzadeh G, Pasbakhsh P, Barbarestani M, Latifpour M - Avicenna J Med Biotechnol (2015 Jul-Sep)

Bottom Line: WJ-MSCs expressed high levels of CD90 (93.6%) and CD105 (90.7%), but low levels of CD34 (0.3%) and CD45 (0.8%).Albumin production had significant difference in the two groups (p≤0.05).The data showed specific characteristics in favor of considering the differentiated cells as hepatocyte-like cells such as obtaining morphologic, functional, and αFP and HNF1-α expression patterns which in turn were higher in cells exposed to RA.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.

ABSTRACT

Background: Wharton's Jelly-Mesenchymal Stem Cells (WJ-MSCs) are pluripotent cells with differentiation capability into most cell lineages. The aim of the current work was to examine the role of Retinoic Acid (RA) in differentiation process of these cells into hepatocyte-like cells and determine the morphological and functional patterns.

Methods: Human WJ-MSCs were extracted, cultured and expanded; after approximately 95% of confluence, the cells were treated with hepatogenic media containing RA. The cells were subsequently analyzed for morphological changes, glycogen storage, albumin production, and specific gene expression.

Results: WJ-MSCs expressed high levels of CD90 (93.6%) and CD105 (90.7%), but low levels of CD34 (0.3%) and CD45 (0.8%). Albumin production had significant difference in the two groups (p≤0.05). The data showed specific characteristics in favor of considering the differentiated cells as hepatocyte-like cells such as obtaining morphologic, functional, and αFP and HNF1-α expression patterns which in turn were higher in cells exposed to RA.

Conclusion: Based on the data of present study, RA is an effective molecule in inducing differentiation of WJ-MSCs into hepatocyte-like cells; therefore, it may be considered as a promising factor for targeting therapy of liver disorders.

No MeSH data available.


Related in: MedlinePlus

Hepatocyte-like cells morphological features. A) Serum free cells show no conspicuous growth alteration, B) short appendage flattened in cells exposed to RA as a contrast with C) not exposed cells after two weeks, D) after three weeks, polyhedral shapes were detected in exposed cells despite of E) not exposed cells, F) at the end of week's four, more visible granules appeared in cells receiving RA than G) not receiving cells, H) morphology of two hexagonal HLCs stained with hematoxylin followed by I) further bile canaliculi-like (arrow) development in exposed cells. All observed with invert microscope. Scale bars for A, B and C=10 μm; for D, E, F, and G=100 μm; for H=20 μm; and for I=50 μm.
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Figure 3: Hepatocyte-like cells morphological features. A) Serum free cells show no conspicuous growth alteration, B) short appendage flattened in cells exposed to RA as a contrast with C) not exposed cells after two weeks, D) after three weeks, polyhedral shapes were detected in exposed cells despite of E) not exposed cells, F) at the end of week's four, more visible granules appeared in cells receiving RA than G) not receiving cells, H) morphology of two hexagonal HLCs stained with hematoxylin followed by I) further bile canaliculi-like (arrow) development in exposed cells. All observed with invert microscope. Scale bars for A, B and C=10 μm; for D, E, F, and G=100 μm; for H=20 μm; and for I=50 μm.

Mentions: The cessation of cell proliferation was observed during serum free time period (Figure 3A). In the presence of RA, cells acquired shortened appendages and broadened and flattened shape (Figure 3B), whereas the changes were not remarkable in cells without exposure to RA after two weeks (Figure 3C). Three weeks after induction, RA led cells into developing polyhedral shapes (Figure 3D) which were in contrast with the shapes of cells in the absence of this factor (Figure 3E). Abundant cytoplasmic granules appeared markedly in exposed cells after four weeks (Figure 3Fvs.3G). Hepatocyte-like cells could obtain hexagonal structure with large nucleus (Figure 3H) and further canalicular architecture (Figure 3I) following prolonged culture in exposed cells.


Retinoic Acid as the Stimulating Factor for Differentiation of Wharton's Jelly-Mesenchymal Stem Cells into Hepatocyte-like Cells.

Mortezaee K, Minaii B, Sabbaghziarani F, Ragerdi Kashani I, Hassanzadeh G, Pasbakhsh P, Barbarestani M, Latifpour M - Avicenna J Med Biotechnol (2015 Jul-Sep)

Hepatocyte-like cells morphological features. A) Serum free cells show no conspicuous growth alteration, B) short appendage flattened in cells exposed to RA as a contrast with C) not exposed cells after two weeks, D) after three weeks, polyhedral shapes were detected in exposed cells despite of E) not exposed cells, F) at the end of week's four, more visible granules appeared in cells receiving RA than G) not receiving cells, H) morphology of two hexagonal HLCs stained with hematoxylin followed by I) further bile canaliculi-like (arrow) development in exposed cells. All observed with invert microscope. Scale bars for A, B and C=10 μm; for D, E, F, and G=100 μm; for H=20 μm; and for I=50 μm.
© Copyright Policy
Related In: Results  -  Collection

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Figure 3: Hepatocyte-like cells morphological features. A) Serum free cells show no conspicuous growth alteration, B) short appendage flattened in cells exposed to RA as a contrast with C) not exposed cells after two weeks, D) after three weeks, polyhedral shapes were detected in exposed cells despite of E) not exposed cells, F) at the end of week's four, more visible granules appeared in cells receiving RA than G) not receiving cells, H) morphology of two hexagonal HLCs stained with hematoxylin followed by I) further bile canaliculi-like (arrow) development in exposed cells. All observed with invert microscope. Scale bars for A, B and C=10 μm; for D, E, F, and G=100 μm; for H=20 μm; and for I=50 μm.
Mentions: The cessation of cell proliferation was observed during serum free time period (Figure 3A). In the presence of RA, cells acquired shortened appendages and broadened and flattened shape (Figure 3B), whereas the changes were not remarkable in cells without exposure to RA after two weeks (Figure 3C). Three weeks after induction, RA led cells into developing polyhedral shapes (Figure 3D) which were in contrast with the shapes of cells in the absence of this factor (Figure 3E). Abundant cytoplasmic granules appeared markedly in exposed cells after four weeks (Figure 3Fvs.3G). Hepatocyte-like cells could obtain hexagonal structure with large nucleus (Figure 3H) and further canalicular architecture (Figure 3I) following prolonged culture in exposed cells.

Bottom Line: WJ-MSCs expressed high levels of CD90 (93.6%) and CD105 (90.7%), but low levels of CD34 (0.3%) and CD45 (0.8%).Albumin production had significant difference in the two groups (p≤0.05).The data showed specific characteristics in favor of considering the differentiated cells as hepatocyte-like cells such as obtaining morphologic, functional, and αFP and HNF1-α expression patterns which in turn were higher in cells exposed to RA.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.

ABSTRACT

Background: Wharton's Jelly-Mesenchymal Stem Cells (WJ-MSCs) are pluripotent cells with differentiation capability into most cell lineages. The aim of the current work was to examine the role of Retinoic Acid (RA) in differentiation process of these cells into hepatocyte-like cells and determine the morphological and functional patterns.

Methods: Human WJ-MSCs were extracted, cultured and expanded; after approximately 95% of confluence, the cells were treated with hepatogenic media containing RA. The cells were subsequently analyzed for morphological changes, glycogen storage, albumin production, and specific gene expression.

Results: WJ-MSCs expressed high levels of CD90 (93.6%) and CD105 (90.7%), but low levels of CD34 (0.3%) and CD45 (0.8%). Albumin production had significant difference in the two groups (p≤0.05). The data showed specific characteristics in favor of considering the differentiated cells as hepatocyte-like cells such as obtaining morphologic, functional, and αFP and HNF1-α expression patterns which in turn were higher in cells exposed to RA.

Conclusion: Based on the data of present study, RA is an effective molecule in inducing differentiation of WJ-MSCs into hepatocyte-like cells; therefore, it may be considered as a promising factor for targeting therapy of liver disorders.

No MeSH data available.


Related in: MedlinePlus