ABSTRACT: Gene array analysis of primary mouse calvaria osteoblasts without differentiation, with differentiation, or with differentiation plus cabozantinib (XL184)
Project description:We performed a gene array analysis using RNAs prepared from undifferentiated, differentiated, or differentiated + cabozantinib (XL184) treated osteoblasts.
Project description:Mucolipidosis type II (MLII) is a severe inherited multisystemic disorder caused by mutations in the GNPTAB gene. Skeletal abnormalities are a predominant feature of MLII. Here we investigate the gene expression in a knock-in mouse model for mucolipidosis type II, generated by the insertion of a cytosine in the murine Gnptab gene (c.3082insC) that is homologous to a homozygous mutation in an MLII patient. Since osteoblasts are critically involved in regulating bone development and remodeling, a genome-wide expression analysis was performed with RNA isolated from primary cultures of osteoblasts originating from MLII knock-in mice (KI) compared to RNA from wild-type (WT) osteoblasts to identify dysregulated genes involved in pathogenic mechanisms. Primary osteoblasts were isolated from calvaria of 5-day-old wild-type (WT) and MLII knock-in littermates (KI). RNA was extracted at day 10 of differentiation induced by ascorbic acid and beta-glycerophosphate and hybridization on Affymetrix microarrays. We used preparations of RNA from two individual primary cultures of osteoblasts for every genotype (WT_OB_I, WT_OB_II, KI_OB_I, KI_OB_II) and compared WT vs KI samples.
Project description:Primary calvaria osteoblasts from newborn mice were cultured. Cells were transfected with control and miR-195 mimic. Following serum starvation cells were treated with recombinant human BMP2 and expression was analyzed 4 hours after treatment. Additionally, osteoblast differentiation was induced with osteogenic medium. Cells were analyzed at confluence (day 0) and 9 days later (day 9).
Project description:Expression data from primary osteoblasts isolated from calvaria of Gnptab(c.3082insC) knock-in mice compared with osteoblasts isolated from wild-type littermates
Project description:Osteoblasts derived from different functional skeletal sites were analysed for gene expression profiles Keywords: Cell type comparison Collagenase digested osteoblasts were derived from two bone sites (calvaria and ulna) of an adult rat, maintained in culture for 3 passages then prepared for RNA analysis
Project description:The effect of parathyroid hormone (PTH) treatment was tested on osteoblasts prepared from calvaria of C57BL/6J mice 7-10 days old. Osteoblasts were treated with either human parathyroid hormone (1-34), bovine (D-Trp12,Tyr34) parathyroid hormone (7-34) amide or vehicle control for 4 hr. Gene expression effects were evaluated by DNA microarray.
