Features include:
- Robust, validated protocol for reprogramming genetically stable EPCs derived from human peripheral or cord blood
- Virus- and DNA-free OKSiM reprogramming factors using self-replicative RNA
- Two-transfection protocol eliminates need for conditioned medium, co-transfections
TABLE 1. Efficient Establishment and Cellular srRNA Reprogramming of EPCs Derived from Human Blood Samples
BLOOD SOURCE | PRIMARY EPC ESTABLISHMENT EFFICIENCY | PATIENT-TO-PATIENT REPROGRAMMING EFFICIENCY |
---|---|---|
Peripheral Blood | 18/22 = 82% | 9/13 = 70% |
Cord Blood | 46/53 = 87% | 33/41 = 81% |
TOTAL | 64/75 = 85% | 42/54 = 78% |
Primary EPC establishment efficiency for peripheral blood is cumulative of both fresh blood and cryopreserved mononuclear cell (MNC) samples. Cord blood efficiency data are from cryopreserved MNCs only. Each collagen-coated T-75 flask was seeded with a minimum of 5 × 107 MNCs. The unmodified StemRNA-SR kit protocol demonstrated high patient-to-patient efficiency using a single reprogramming attempt. Typical final yields are 10-20 iPS cell colonies per well in a 6-well plate format.