Generation of iPS cell line
INGESTEM has the widest collection of iPSC cell lines produced in France. More than 200 normal or pathological human iPSC lines are available and distributed for research projects (www.hescreg.eu).
INGESTEM provides the academic and industrial community technology to generate iPS cell lines on demand, from primary cells with genotypes of interest.
Three services are available:
- iPSC production for exclusive use (on-demand)
- Production of iPSC line “research tool” in the framework of a collaborative work
- Maintenance in culture of your iPSC lines (annual subscription)
Key steps
Preparation of somatic cells
- Administrative preparation
- Selection of donor, collection of biological and genetic data
- Sanitary tests on cells
- Creation of a master cell bank
cellular reprogramming
- Culture of somatic cells
- Reprogramming by non-integrative methods
Selection, characterization and amplification of iPSC clones
- Selection of 4 iPS clones by morphology and pluripotency markers.
- Genomic integrity tests and pluripotency assessment in vivo
- Amplification of a primary bank of iPS line (clones and polyclonal cell lines)
- Viability test after freeze / thaw cycles.
Upstream of the project, a contrat and Material Transfert Agreement are prepared, including the ethical issues (Informed consent, ethics committee or IRB ). Samples are anonymized.
Many types of primary cells can be reprogrammed iPSC: mononuclear blood cells, fibroblasts, mesenchymal cells, cord blood cells, epithelial cells, etc.
The sanitary status of primary cells is a pre-request before reprogramming (mycoplasma, HIV, CMV, HCV, HBV) as well as the genotype (karyotype, CNV, SNP, mutation).
Cells are reprogrammed exclusively by a non-integrative methods: Sendai virus, mRNA or episomes.
