iPSC Generation from Patient Samples
Our scientists are experts in the generation of induced pluripotent stem cell (iPSC) lines from patient specific fibroblasts or blood cells. We have three methods for iPSC generation: mRNA, episomal (both genomic footprint-free), or retroviral.
Genomic footprint-free iPSC Generation Service - our custom cell line generation service.
No genomic footprint - the non-integrating system is safe in drug discovery and cell therapy applications.
Two methods available: Episomal DNA vectors containing the reprogramming factors or mRNA.
High reprogramming efficiency (0.2%-0.5%).
The reprogramming vectors/mRNA are removed naturally during the cell cycle.
Safe to handle - no viral particles.
You will need to provide: 1 x 106 cells from patient fibroblasts or PBMC cells from your patient samples.
- At least 3 clones with two vials of each clone (>2 x 105 viable cells/ vial)
- Clones are characterized by pluripotency markers (OCT4, SOX2, SSEA4, TRA-1-60, TRA-1-80)
- Embryoid body formation (optional)
- Teratoma analysis (optional)
Applied StemCell, Inc. can create CRISPR/Cas9 modified iPSCs using patient samples.
CASE STUDY – Patient derived iPSC Generation
ASC has successfully generated iPSCs from several disease specific patient samples. Here is one such project:
Case Study: Generation of Human iPSCs from Patient-derived Skin Fibroblasts
Goal: To generate an iPSC line from patient derived dermal fibroblasts.
The dermal fibroblasts were obtained by skin biopsy from a patient with a rare disorder. The fibroblasts were plated, transfected via electroporation and cultured with ASC’s proprietary iPSC generation media until iPSC colonies appeared. Five high quality candidate colonies were expanded and characterized by immunocytochemical staining using primary antibodies for: OCT4, SOX2, SSEA4, TRA-1-60 and TRA-1-81) and alkaline phosphatase staining. All five clones stained positive for various pluripotency markers tests as well as for alkaline phosphatase (Figure1).
Figure 1. Representative iPSC clones reprogrammed from patient derived fibroblasts. The clones tested positive for pluripotency markers: OCT4, SOX2, SSEA4, TRA-160, TRA-1-81 and for alkaline phosphatase (AP).
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