Cell culture of p63 knock out and wild type bladder epithelial stem cell. Mus musculus

Although there are several reports describing a stem cell fraction to form urothelium and lineage tracing studies in urinary bladder, it is still unclear about stem cells in mouse bladder epithelium. Here, we have established the cloning system of mouse stem cells from wild type and p63 knockout bladder. We isolated stem/progenitor cells from wild type and p63 knockout epithelium. From wild type bladder, we have cloned keratin 5-positive stem cells, and from p63 knockout epithelium, which loses basal cell fraction, we have cloned morphologically-distinct keratin 8 and claudin 3 positive stem cells, which may be reminiscent of intestinal metaplasia in bladder. Overall design: From newborn mice (p63 knockout and wild type), bladder were collected and digested with collagenase for stem cell culture. Epithelial stem cells were cultured on 3T3 feeder cells. Colonies of epithelial stem cells from p63 knockout and wild type bladder were picked up, and total RNA was isolated with Picopure RNA isolation kit. We used affymetrix mouse exon 1.0 st array to hybridize 2 samples from each of tissues (p63 knockout and wild types).