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Effect of acute hypoxia on CXCR4 gene expression in C57BL/6 mouse bone marrow-derived mesenchymal stem cells.

Kadivar M, Alijani N, Farahmandfar M, Rahmati S, Ghahhari NM, Mahdian R - Adv Biomed Res (2014)

Bottom Line: Moreover, the relative gene expression of CXCR4 was decreased after hypoxia-reoxygenation by more than 80% in 4 h (0.136 ± 0.018) and 24 h (12.77 ± 0.707) groups.The results suggest that CXCR4 expression in MSCs decreases upon acute hypoxic stress.This difference could have resulted from the cells being compatible with low oxygen metabolism.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Pasteur Institute of Iran, Tehran, Iran.

ABSTRACT

Background: One of the most important stimuli in stem cell biology is oxygen. Chemokine receptor 4 (CXCR4) plays a crucial role in the migration and homing of stem cells. In this study, mesenchymal stem cells (MSCs) were exposed to 1% oxygen to investigate the effect of acute hypoxia on CXCR4 gene expression.

Materials and methods: MSCs were isolated from C57BL/6 mouse bone marrow and were identified and expanded in normoxic culture. Cells were incubated at 37°C under 1% hypoxic conditions for periods of 4, 8, 16, 24, and 48 h. After hypoxia preconditioning, the cells were placed in normoxic condition for 8 h to achieve cellular hypoxia-reoxygenation. To assess the level of CXCR4 gene expression, real-time quantitative reverse transcription-polymerase chain reaction was carried out for each group.

Results: Data from statistical analysis illustrated that exposure of MSCs to acute hypoxic condition down-regulates CXCR4 expression with the maximum under-expression observed in 4 h (0.91 ± 0.107) and 8 h (50 ± 2.98) groups. Moreover, the relative gene expression of CXCR4 was decreased after hypoxia-reoxygenation by more than 80% in 4 h (0.136 ± 0.018) and 24 h (12.77 ± 0.707) groups.

Conclusion: The results suggest that CXCR4 expression in MSCs decreases upon acute hypoxic stress. Furthermore, hypoxia-reoxygenated MSCs showed decreased expression of CXCR4, compared to cells subjected to acute hypoxia. This difference could have resulted from the cells being compatible with low oxygen metabolism. In summary, before the therapeutic application of MSCs, it should be regarded as a necessity to optimize the oxygen concentration in these cells, as it is a critical factor in modulating CXCR4 expression.

No MeSH data available.


Related in: MedlinePlus

Adipogenic differentiation of bone marrow-derived mesenchymal stem cells. Oil Red O staining of cultures that were treated (a) and not treated (b) with adipogenic media for 3 weeks
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Figure 2: Adipogenic differentiation of bone marrow-derived mesenchymal stem cells. Oil Red O staining of cultures that were treated (a) and not treated (b) with adipogenic media for 3 weeks

Mentions: MSCs isolated from C57BL/6 mice bone marrow were cultured in appropriate medium as described and reached 80% confluence by day 14. Cells at the second passage stained strongly for CD73 and STRO-1. Positive staining for these markers confirmed that the isolated cells were MSCs [Figure 1]. To verify that immunodepleted MSCs were multipotent, their ability to differentiate into osteocytes and adipocytes was analyzed. After 10 days in osteogenic medium or 21 days in adipogenic medium, MSCs differentiated into adipocytes [Figure 2] or osteocytes [Figure 3] as indicated by increase of Oil Red O or alizarin red staining, respectively.


Effect of acute hypoxia on CXCR4 gene expression in C57BL/6 mouse bone marrow-derived mesenchymal stem cells.

Kadivar M, Alijani N, Farahmandfar M, Rahmati S, Ghahhari NM, Mahdian R - Adv Biomed Res (2014)

Adipogenic differentiation of bone marrow-derived mesenchymal stem cells. Oil Red O staining of cultures that were treated (a) and not treated (b) with adipogenic media for 3 weeks
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4260281&req=5

Figure 2: Adipogenic differentiation of bone marrow-derived mesenchymal stem cells. Oil Red O staining of cultures that were treated (a) and not treated (b) with adipogenic media for 3 weeks
Mentions: MSCs isolated from C57BL/6 mice bone marrow were cultured in appropriate medium as described and reached 80% confluence by day 14. Cells at the second passage stained strongly for CD73 and STRO-1. Positive staining for these markers confirmed that the isolated cells were MSCs [Figure 1]. To verify that immunodepleted MSCs were multipotent, their ability to differentiate into osteocytes and adipocytes was analyzed. After 10 days in osteogenic medium or 21 days in adipogenic medium, MSCs differentiated into adipocytes [Figure 2] or osteocytes [Figure 3] as indicated by increase of Oil Red O or alizarin red staining, respectively.

Bottom Line: Moreover, the relative gene expression of CXCR4 was decreased after hypoxia-reoxygenation by more than 80% in 4 h (0.136 ± 0.018) and 24 h (12.77 ± 0.707) groups.The results suggest that CXCR4 expression in MSCs decreases upon acute hypoxic stress.This difference could have resulted from the cells being compatible with low oxygen metabolism.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Pasteur Institute of Iran, Tehran, Iran.

ABSTRACT

Background: One of the most important stimuli in stem cell biology is oxygen. Chemokine receptor 4 (CXCR4) plays a crucial role in the migration and homing of stem cells. In this study, mesenchymal stem cells (MSCs) were exposed to 1% oxygen to investigate the effect of acute hypoxia on CXCR4 gene expression.

Materials and methods: MSCs were isolated from C57BL/6 mouse bone marrow and were identified and expanded in normoxic culture. Cells were incubated at 37°C under 1% hypoxic conditions for periods of 4, 8, 16, 24, and 48 h. After hypoxia preconditioning, the cells were placed in normoxic condition for 8 h to achieve cellular hypoxia-reoxygenation. To assess the level of CXCR4 gene expression, real-time quantitative reverse transcription-polymerase chain reaction was carried out for each group.

Results: Data from statistical analysis illustrated that exposure of MSCs to acute hypoxic condition down-regulates CXCR4 expression with the maximum under-expression observed in 4 h (0.91 ± 0.107) and 8 h (50 ± 2.98) groups. Moreover, the relative gene expression of CXCR4 was decreased after hypoxia-reoxygenation by more than 80% in 4 h (0.136 ± 0.018) and 24 h (12.77 ± 0.707) groups.

Conclusion: The results suggest that CXCR4 expression in MSCs decreases upon acute hypoxic stress. Furthermore, hypoxia-reoxygenated MSCs showed decreased expression of CXCR4, compared to cells subjected to acute hypoxia. This difference could have resulted from the cells being compatible with low oxygen metabolism. In summary, before the therapeutic application of MSCs, it should be regarded as a necessity to optimize the oxygen concentration in these cells, as it is a critical factor in modulating CXCR4 expression.

No MeSH data available.


Related in: MedlinePlus