The core reprograms fibroblasts and blood cells (peripheral blood mononuclear cells) into iPSCs using Sendai virus. The process of reprogramming usually takes 2-4 months and includes expansion of fibroblasts or blood cells, iPSC colony picking, and expansion of iPSC colonies.
iPSCs are cultured, incubated, and expanded on Matrigel covered wells with Essential 8. Our facility has incubators and biological safety cabinets for the culturing of iPSCs.
We use published protocols to differentiate our iPSCs into cardiomyocytes, fibroblasts, endothelial cells, and neurons. Differentiation can take between 10-30 days, depending on the protocol. Contact us for more details on the type of cells you are interested in generating from iPSCs.
Banking and Distribution
We offer banking services, in which we can freeze, thaw, expand, and recover iPSC lines. iPSC lines and cell lineages derived from iPSCs are cryopreserved in vials and distributed to other institutes.
We perform quality control on all iPSC lines. Lines are routinely tested for mycoplasma contamination using PCR. We also perform karyotype analysis and SNP arrays to authenticate the lines being distributed.
The core performs immunofluorescence staining to confirm the expression of pluripotency markers. The iPSCs are stained using antibodies specific to the markers and observed on a microscope.
We use qRT-PCR to determine the levels of pluripotency markers such as Oct3/4, Nanog, and Sox2 and to characterize the progress of differentiation. This can determine if lines are able to be differentiated and how well they will respond to differentiation protocols.
Virtual karyotyping is performed to determine the level of genomic stability in iPSCs. We use SNP microarray data of the cell lines of interest and compare it to a database of similar expression patterns to find genomic abnormalities.
We offer hands-on training on the basics of iPSCs. Training includes how to culture, pass, freeze, thaw, and differentiate iPSCs.