Reprogramming
We offer reprogramming of somatic patient-derived cells to establish stable iPSC lines which are subsequently robustly characterized and banked for the researcher.
Currently we offer reprogramming of fibroblasts (from skin) and PBMCs.
We have also established reprogramming protocols for other somatic cell types (incl. urinary cells and T-cells). If you are interested in this, please contact us.
Primary cells can be provided to us in culture or cryopreserved. Additionally we offer the isolation of primary cells:
- PBMC from whole blood samples using CPT tubes
- Fibroblasts from skin biopsies
We only apply reprogramming methods that utilize non-integrating vectors. PBMC will be reprogrammed using sendai viral vectors, Fibroblasts will be reprogrammed by self-replicating RNA or sendai viral vectors.
For details on the reprogramming workflow see section below.
Ethics
Before starting any project involving hiPSC generation from patients a project specific vote by the Ethikkommission of the Charité is mandatory. Here hiPSC specific details in the patient information and consent forms are important. To support these applications we offer advice and template forms that can be downloaded in the Resource section.
Reprogramming workflow
The reprogramming process in total takes about three month from primary cell isolation until clones are ready for banking and quality control. From each donor sample we isolate five clones and freeze down seed banks. Based on quality control criteria (including CNV analysis using SNP arrays and culture characteristics) one clone will be selected and subjected to banking, characterization and quality control.
