Evaluation of two cell culture media in culturing rat full term amniotic fluid cells

Introduction: Amniotic fluid (AF) consists of heterogenous population of cells with high diagnostic and therapeutic values. The study of rat amniotic fluid cells is very limited, despite the extensive use of this animal model in biomedical research. Primary culture of rat AF cells, especially from full term pregnancies has not been well established. Here we attempt to determine the suitable medium in culturing rat AF cells that would enhance the cell viability, growth rate and heterogeneity. Methods: The cell viability, growth rate and heterogeneity of rat AF cells were compared upon culturing the primary cells in two different media; Amniomax or RPMI. Cell viability study was carried out using trypan blue staining, while the growth rate was monitored based on the time required to passage the cells (population doubling time in hour). The heterogeneity of cells was examined based on the morphology of the cells. Statistical analysis was performed using t-test. Results: Amniomax was observed to provide a better culture condition in culturing rat AF cells as the cells are more viable, grow faster and more heterogenous as compared to the cells grown in RPMI. Conclusion: Amniomax is a more suitable medium for high quality and viability of full term rat AF cell culture, as compared to RPMI. Thus, warranting propagation of more rat AF cells for biomedical research

Downregulation of RhoGDIα increased migration and invasion of ER+ MCF7 and ER− MDA-MB-231 breast cancer cells.

Rho GDp dissociation inhibitors (RhoGDIs) can inhibit cell motility, invasion, and metastasis in cancer by inactivating the RhoGTpases. A member of RhoGDI family has been consistently shown to interact with estrogen receptor (eR), and change its transcriptional activity. eR is a receptor known to be inversely correlated with cell motility and invasion in breast cancer. The consequence of RhoGDIα activity on migration and invasion of eR+ and eR− breast cancers is not clear. The aim of our study was to investigate the possible opposing effect of RhoGDIα on the migration and invasion of eR+ MCF7 and eR− MDA-MB-231 breast cancer cells. RhoGDIα was downregulated using short interfering RNA (siRNA) and upregulated using GFp-tagged ORF clone of RhoGDIα, and their ability for migration and invasion was assayed using transwell chambers. It was found that the silencing of RhoGDIα in MCF7 and MDA-MB-231 cells significantly increased migration and invasion of these cells into the lower surface of porous membrane of the chambers. Overexpression of RhoGDIα in MCF7 cells suppressed their migration and invasion, but no significant effect was found on MDA-MB-231 cells. Our results indicate that the downregulation of RhoGDIα similarly affects the in vitro migration and invasion of eR+ MCF7 and eR− MDA-MB-231 cells. however, our assays are differently affected by the upregulation of RhoGDIα in these two cell lines and this may be due to the differences in eR expression, primary invasive ability and/or other molecules between these two cell line models which warrant further investigation

Phosphate homeostasis and genetic mutations of familial hypophosphatemic rickets

Hypophosphatemic rickets (HR) is a syndrome of hypophosphatemia and rickets that resembles vitamin D deficiency, which is caused by malfunction of renal tubules in phosphate reabsorption. Phosphate is an essential mineral, which is important for bone and tooth structure. It is regulated by parathyroid hormone, 1,25-dihydroxyvitamin D and fibroblast-growth-factor 23 (FGF23). X-linked hypophosphatemia (XLH), autosomal dominant HR (ADHR), and autosomal recessive HR (ARHR) are examples of hereditary forms of HR, which are mainly caused by mutations in the phosphate regulating endopeptidase homolog, X-linked (PHEX), FGF23, and, dentin matrix protein-1 (DMP1) and ecto-nucleotide pyro phosphatase/phosphodiesterase 1 (ENPP1) genes, respectively. Mutations in these genes are believed to cause elevation of circulating FGF23 protein. Increase in FGF23 disrupts phosphate homeostasis, leading to HR. This review aims to summarize phosphate homeostasis and focuses on the genes and mutations related to XLH, ADHR, and ARHR. A compilation of XLH mutation hotspots based on the PHEX gene database and mutations found in the FGF23, DMP1, and ENPP1 genes are also made available in this review

Preterm intraamniotic infection and inflammation: search for protein biomarkers via proteomics approach

Preterm delivery (PTD) is commonly caused by intraamniotic infection and inflammation (IAI) and is one of the leading causes of maternal and neonatal morbidity or mortality worldwide. IAI in pregnant women with high risk of PTD can be classified according to the membrane status; preterm labour with intact membranes (PLIM) and preterm pre-labour rupture of membranes (PPROM). At present, the treatment for PTD-associated IAI is principally based on antibiotic therapy. However, this therapy is found to be only beneficial to PPROM. Several IAI related protein biomarkers have been identified. Unfortunately, most of the studies that discovered the biomarkers focused on categorising term delivery and PTD-associated IAI, and to a lesser extent on the membrane status. Therefore, identifying a panel of highly sensitive and specific protein biomarkers that are able to discriminate IAI pregnant women at high risk of PTD with PLIM from PPROM are urgently needed to improve the treatment benefits. Over the past decade, proteomics technologies have been applied widely in identifying PTD-associated IAI biomarkers. This review brings together information on PTD-associated IAI protein biomarkers and touches on the combination of multivariable biomarkers to develop a more sensitive and specific biomarkers to discriminate PTD-associated IAI with PLIM from PPROM. Proteomics technologies and its workflow on protein biomarkers discovery are also included in this review