Human iPSC-derived lipids
Human cardiomyocyte lipids and CNS lipids are extracted from Ncardia's
Cor.4U® cardiomyocytes and CNS.4U® neural cells, respectively. Both cell lines are produced by in vitro differentiation of transgenic human induced pluripotent stem cells(iPSC), generated by introducing defined transcription factors, as described by Yamanaka et al in human skin fibroblasts, using a non-viral system, and are grown in fully synthetic media. Cardiomyocytes and CNS cells are well-characterized and of high purity, and their human origin ensures optimal composition for use in human membrane protein assays.
Human iPSC-derived lipids can restore membrane protein functionality in artificial surroundings:
SPR and other ligand-binding assays (Figure 1)
Detailed protocols for successful purification
The highly pure cardiomyocytes express cardiac-specific proteins, e.g., cardiac alpha-actinin and the gap junction protein connexin-43.
CNS cells comprise key cell types of the human central nervous system (CNS): neurons (80-90%) and astrocytes (10-20%). Of the neurons, 90% are glutamatergic and GABAergic and 10% are dopaminergic neurons.
Figure 1: Human CNS lipids restore membrane protein functionality in surface plasmon resonance (SPR). SPR measurements were performed by immobilization of target proteins via a His tag on a Ni-NTA-functionalized chip and monitoring association/dissociation kinetics of the specific ligand Taurolithocholic acid. A: GPBAR1/TGR5 membrane protein preparation in detergent, incubated in the presence of Human CNS lipids_iPSC-derived o/n at 4°C; binding constants of 11 nM were measured, which is in accordance with published values. B: The same protein preparation was incubated at identical conditions, but without any Human CNS lipids added. No signal was obtained in this measurement.
1. Takahashi, K et al. (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell Nov 30;131(5):861-72
2. Axiogenesis AG Patent WO2012098260 A1, A non-viral system for generation of induced pluripotent stem (iPS) cells