iPSC Generation

mRNA iPSC Generation

Induced Pluripotent Stem Cells

Induced Pluripotent Stem Cell Generation: our mRNA iPSC generation method uses a transgene-free, footprint-free iPSC generation with our non-integrating transfection system. This provides safety for the iPSC to be used in drug discovery and cell therapy applications.  Molecular and functional assays such as flow cytometry, karyotyping, and in-vitro Embryoid body formation is used to characterize the pluripotency of the newly established iPSC cell line.

Skin Fibroblasts

mRNA Reprogramming

iPSC Generation

mRNA iPSC Reprogramming

Our method of cellular reprogramming of skin cells to Induced Pluripotent Stem Cells uses a custom mRNA method. This means we ARE NOT using viruses or episomal methods to induced pluripotency. Our protocol generates CLEAN iPSCs which can be used as a source to induced differentiated cell types for therapeutic applications.

The iPSC21 protocol requires on average 2 months to cultures the skin fibroblasts (starting cell type), reprogram the cells with our mRNA method, subculture and expand the generated iPSCs, then finally the validation of iPSCs (some parameters are shown below). At this stage, the iPSCs are cryogenically preserved for future use.

All iPSC21 customers will get a full certification of analysis (COA)

iPSC Characterisation

Validation of Pluipotency

Immunocytochemistry Staining (ICC)

Alkaline phosphatase (AP) is a universal pluripotent marker for all types of pluripotent stem cells including embryonic stem cells, embryonic germ cells, and induced pluripotent stem cells. The pluripotent status of stem cells can be characterized by a high level of AP expression, along with the expression of multiple pluripotency markers including the transcription factors Nanog, Oct4, Sox2, stage-specific embryonic antigens, SSEA-1, -3, -4, and tumor-related antigens, TRA-1-60, TRA-1-81.

Alkaline Phosphatase (AP)

OCT-4

SOX-2

TRA-1-60

SSEA-4

DAPI

iPSC Phenotype by Flow Cytometry

Flow cytometry, utilizing antibodies recognizing targets restricted to the iPSC surfaceome, offers an invaluable tool for high-throughput validation of iPSC lines.

Part of the SC21 Group