iPSC Quality Check
We offer quality control (QC) services for newly generated lines or imported lines of interest. Recommended QC based on ISSCR guidelines include: line hygiene, pluripotency, potency and genomic stability.
Line pluripotency assessment is based on morphology (defined colony boundaries, compact colonies and minimal spontaneous differentiation) and expression of key pluripotency factors. Pluripotency factors can be assessed by immunofluorescence or flow cytometry.
The potency of iPSC lines will be assessed by directed differentiation towards ectoderm (Pax6 Sox2 double positive cells) , mesoderm (CD144 and CD140b single and double positive cell populations) and endoderm (Sox17 and CxcR4 double positive cell population). Differentiation efficiency will be assessed by flow cytometry using a cut-off of 50% commitment to all lineages per clone using non-optimised seeding protocols.
Assessment of genomic stability is critical at the level of your laboratory master cell bank and when considering a specific iPSC line for gene editing. In the case of editing, it is also important to check the resulting single cell edited clones against the parental line to ensure that the monoclonalisation process has not resulted in the acquisition of SNPs that confer a proliferative advantage and a differentiation bias to these clones. We only offer karyotyping based genotyping using the Global Screening Array (GSA) v4.0 as a collaborative effort between our core and Core Facility Genomics (CF-GEN).
Our services aim to ensure the basic quality required for reproducible use of iPS cultures for maintenance and differentiation.
We offer quality control (QC) services for newly generated lines or imported lines of interest. Recommended QC based on ISSCR guidelines include: line hygiene, pluripotency, potency and genomic stability.
Line pluripotency assessment is based on morphology (defined colony boundaries, compact colonies and minimal spontaneous differentiation) and expression of key pluripotency factors. Pluripotency factors can be assessed by immunofluorescence or flow cytometry.
The potency of iPSC lines will be assessed by directed differentiation towards ectoderm (Pax6 Sox2 double positive cells) , mesoderm (CD144 and CD140b single and double positive cell populations) and endoderm (Sox17 and CxcR4 double positive cell population). Differentiation efficiency will be assessed by flow cytometry using a cut-off of 50% commitment to all lineages per clone using non-optimised seeding protocols.
Assessment of genomic stability is critical at the level of your laboratory master cell bank and when considering a specific iPSC line for gene editing. In the case of editing, it is also important to check the resulting single cell edited clones against the parental line to ensure that the monoclonalisation process has not resulted in the acquisition of SNPs that confer a proliferative advantage and a differentiation bias to these clones. We only offer karyotyping based genotyping using the Global Screening Array (GSA) v4.0 as a collaborative effort between our core and Core Facility Genomics (CF-GEN).
Our services aim to ensure the basic quality required for reproducible use of iPS cultures for maintenance and differentiation.
