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Non-integrating episomal reprogramming. Reprogramming and quality control performed by Cellular Dynamics International. Donors screened and negative for HIV, HBV, and HCV*.
Handling of lines:
*Lines are grown feeder free on E8/vitronectin and are frozen in colony form.
Donor Consenting:Each donors consent included statements on the following:
No deletions or amplifications larger than 5MB (resolution of traditional G-banding assay) on SNP arrays with LogRDev score less than 0.5.
Donor DNA is tested on all abnormal clones to whether the genetic abnormality is due to
|Pluripotency||Analysis of gene expression by qPCR
of 48 mRNAs.
|A non-probabilistic binary linear classifier identifies the gene expression of the sample as
iPSC based on an appropriate training set. This method has been benchedmarked against traditional teratoma assays for pluripotency.
|Genotyping using PCR assay for 48 SNPs.||≤1 mismatch between donor and iPSC line.|
|Endpoint multiplex PCR for 2 plasmid
sequences (EBNA and OriP).
|Detection of ≤1 plasmid copy per 100 cells or a decrease in the number of plasmid copies detected at passage 5.|
|Mycoplasma||qPCR for 8 species.||Negative for all 8 species.|
|Sterility||Microbiological testing by third-‐party
|Negative for all tests.|