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Differentiation analyses of adult suspension mononucleated peripheral blood cells of Mus musculus.

Ariffin SH, Abidin IZ, Yazid MD, Wahab RM - Cell Commun. Signal (2010)

Bottom Line: Differentiation into osteoblast cells was concomitant with the activation of the Opn gene, increment of alkaline phosphatase (ALP) activity and the existence of bone nodules, whereas osteoclast cells activated the Catk gene, increment of tartrate resistant acid phosphatase (TRAP) activity and showed resorption activities via resorption pits.Morphology analyses showed the morphology of osteoblast and osteoclast cells after von Kossa and May-Grunwald-Giemsa staining respectively.In conclusion, suspension mononucleated cells have the potentiality to differentiate into mature osteoblasts and osteoclasts, and hence can be categorized as multipotent stem cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, Malaysia. shahroy8@gmail.com.

ABSTRACT

Background: The purpose of this study is to determine whether isolated suspension mouse peripheral mononucleated blood cells have the potential to differentiate into two distinct types of cells, i.e., osteoblasts and osteoclasts.

Results: Differentiation into osteoblast cells was concomitant with the activation of the Opn gene, increment of alkaline phosphatase (ALP) activity and the existence of bone nodules, whereas osteoclast cells activated the Catk gene, increment of tartrate resistant acid phosphatase (TRAP) activity and showed resorption activities via resorption pits. Morphology analyses showed the morphology of osteoblast and osteoclast cells after von Kossa and May-Grunwald-Giemsa staining respectively.

Conclusions: In conclusion, suspension mononucleated cells have the potentiality to differentiate into mature osteoblasts and osteoclasts, and hence can be categorized as multipotent stem cells.

No MeSH data available.


Related in: MedlinePlus

Expression of specific gene markers. RT-PCR analysis was performed using total RNA isolated from mononucleated cells cultured in proliferation medium and mononucleated cells cultured in osteoblast and osteoclast differentiation medium. (A) Gapdh (717 bp) was used as a positive control. (B) The low expression of Opn (234 bp) in undifferentiated mononucleated cells while high expression of Opn (234 bp) indicates differentiation into mature osteoblasts. (C) Catk (350 bp) expression shows osteoclast differentiation. Equal volumes of the products were separated in agarose gels and stained with ethidium bromide.
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Figure 1: Expression of specific gene markers. RT-PCR analysis was performed using total RNA isolated from mononucleated cells cultured in proliferation medium and mononucleated cells cultured in osteoblast and osteoclast differentiation medium. (A) Gapdh (717 bp) was used as a positive control. (B) The low expression of Opn (234 bp) in undifferentiated mononucleated cells while high expression of Opn (234 bp) indicates differentiation into mature osteoblasts. (C) Catk (350 bp) expression shows osteoclast differentiation. Equal volumes of the products were separated in agarose gels and stained with ethidium bromide.

Mentions: The activation of the Opn gene shows that mononucleated cells have differentiated into osteoblast cells. RT-PCR analysis showed different levels of Opn gene expression in mononucleated cells cultured in proliferation medium compared to osteoblast differentiation medium (Figure 1B). Both media produced the expected products; i.e., the Opn gene with size ~234 bp when amplified with RT-PCR. However, the expression of the Opn gene for mononucleated cells cultured in osteoblast differentiation medium were much higher as compared to mononucleated cells cultured in proliferation medium (Figure 1B). The increase in Opn gene transcription showed that peripheral blood mononucleated cells differentiate into mature osteoblast cells. Opn gene can produce low grade expression in variety of cells, such as fibroblast, differentiated osteoblast and osteoclast cells and bone marrow derived cells [13]. Low level expression of Opn gene in this study might be due to the existence a few of all these differentiated cells types and also primitive cells that can survive after 15 days of medium selection. However, the increment of expression level after been induced in specialized osteoblast medium was contributed by the primitive cell (stem cells) that have been differentiated into osteoblast cells.


Differentiation analyses of adult suspension mononucleated peripheral blood cells of Mus musculus.

Ariffin SH, Abidin IZ, Yazid MD, Wahab RM - Cell Commun. Signal (2010)

Expression of specific gene markers. RT-PCR analysis was performed using total RNA isolated from mononucleated cells cultured in proliferation medium and mononucleated cells cultured in osteoblast and osteoclast differentiation medium. (A) Gapdh (717 bp) was used as a positive control. (B) The low expression of Opn (234 bp) in undifferentiated mononucleated cells while high expression of Opn (234 bp) indicates differentiation into mature osteoblasts. (C) Catk (350 bp) expression shows osteoclast differentiation. Equal volumes of the products were separated in agarose gels and stained with ethidium bromide.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2984458&req=5

Figure 1: Expression of specific gene markers. RT-PCR analysis was performed using total RNA isolated from mononucleated cells cultured in proliferation medium and mononucleated cells cultured in osteoblast and osteoclast differentiation medium. (A) Gapdh (717 bp) was used as a positive control. (B) The low expression of Opn (234 bp) in undifferentiated mononucleated cells while high expression of Opn (234 bp) indicates differentiation into mature osteoblasts. (C) Catk (350 bp) expression shows osteoclast differentiation. Equal volumes of the products were separated in agarose gels and stained with ethidium bromide.
Mentions: The activation of the Opn gene shows that mononucleated cells have differentiated into osteoblast cells. RT-PCR analysis showed different levels of Opn gene expression in mononucleated cells cultured in proliferation medium compared to osteoblast differentiation medium (Figure 1B). Both media produced the expected products; i.e., the Opn gene with size ~234 bp when amplified with RT-PCR. However, the expression of the Opn gene for mononucleated cells cultured in osteoblast differentiation medium were much higher as compared to mononucleated cells cultured in proliferation medium (Figure 1B). The increase in Opn gene transcription showed that peripheral blood mononucleated cells differentiate into mature osteoblast cells. Opn gene can produce low grade expression in variety of cells, such as fibroblast, differentiated osteoblast and osteoclast cells and bone marrow derived cells [13]. Low level expression of Opn gene in this study might be due to the existence a few of all these differentiated cells types and also primitive cells that can survive after 15 days of medium selection. However, the increment of expression level after been induced in specialized osteoblast medium was contributed by the primitive cell (stem cells) that have been differentiated into osteoblast cells.

Bottom Line: Differentiation into osteoblast cells was concomitant with the activation of the Opn gene, increment of alkaline phosphatase (ALP) activity and the existence of bone nodules, whereas osteoclast cells activated the Catk gene, increment of tartrate resistant acid phosphatase (TRAP) activity and showed resorption activities via resorption pits.Morphology analyses showed the morphology of osteoblast and osteoclast cells after von Kossa and May-Grunwald-Giemsa staining respectively.In conclusion, suspension mononucleated cells have the potentiality to differentiate into mature osteoblasts and osteoclasts, and hence can be categorized as multipotent stem cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, Malaysia. shahroy8@gmail.com.

ABSTRACT

Background: The purpose of this study is to determine whether isolated suspension mouse peripheral mononucleated blood cells have the potential to differentiate into two distinct types of cells, i.e., osteoblasts and osteoclasts.

Results: Differentiation into osteoblast cells was concomitant with the activation of the Opn gene, increment of alkaline phosphatase (ALP) activity and the existence of bone nodules, whereas osteoclast cells activated the Catk gene, increment of tartrate resistant acid phosphatase (TRAP) activity and showed resorption activities via resorption pits. Morphology analyses showed the morphology of osteoblast and osteoclast cells after von Kossa and May-Grunwald-Giemsa staining respectively.

Conclusions: In conclusion, suspension mononucleated cells have the potentiality to differentiate into mature osteoblasts and osteoclasts, and hence can be categorized as multipotent stem cells.

No MeSH data available.


Related in: MedlinePlus