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The effects of vitamin D3 (1α,25-dihydroxyvitamin D3) on the haematopoietic differentiation of stem cells in murine in vitro studies and human in vivo studies

Al Qaisi, Mayada (2015) The effects of vitamin D3 (1α,25-dihydroxyvitamin D3) on the haematopoietic differentiation of stem cells in murine in vitro studies and human in vivo studies. Doctoral thesis (PhD), Manchester Metropolitan University.

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Abstract

Previous research using cancer cells has demonstrated that 1α,25-dihydroxyvitamin D3 (VD3) can suppress cell proliferation and promote cell differentiation. The research reported in this research ascertains that VD3 plays a similar role in haematopoiesis. This was investigated using the E14/ OP9 co-culture as it has previously been shown to be a good setting for studying haematopiesis. The effects of VD3 on E14 and OP9 were tested separately with different VD3 levels to demonstrate that sufficient levels of VD3 were crucial in promoting differentiation and suppressing proliferation of both cell lines. The effective dosage of VD3 was 100 nmol/L. This dosage was then used to test the effect of VD3 on cell proliferation and differentiation in haematopoiesis in the E14 / OP9 co-culture system through analyses of growth factors, cluster of differentiation (CD) markers, colony-forming cell (CFC) counts, haematopoiesis-specific genes and cytokine expression. VD3 was shown to promote differentiation thereby accelerating the early appearance of differentiated cells in the co-culture system. The in vitro study was followed by an in vivo pilot study, in which blood samples were taken monthly from a group of adult volunteers over a three-month period for analysis of VD levels, CD34 expression and blood cell counts. In the in vivo pilot study, it was found that there were significant non-random temporal variations in the measurements and there were statistically significant positive correlations between each of 25-hydroxyvitamin D (25(OH)D) level, white blood cell count, platelet count, CD34+ progenitor cell count and VD3 level. Overall, the research reported in this thesis demonstrated for the first time an anabolic effect of VD3 in haematopoiesis on murine E14 and OP9 cells in vitro. In addition, the pilot study on human volunteers demonstrated that a comparable effect takes place on human cells in vivo and these results could be used to inform future studies.

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