Definitive Endoderm Formation from Plucked Human Hair-Derived Induced Pluripotent Stem Cells and SK Channel Regulation

Abstract

Pluripotent stem cells present an extraordinary powerful tool to investigate embryonic development in humans. Essentially, they provide a unique platform for dissecting the distinct mechanisms underlying pluripotency and subsequent lineage commitment. Modest information currently exists about the expression and the role of ion channels during human embryogenesis, organ development, and cell fate determination. Of note, small and intermediate conductance, calcium-activated potassium channels have been reported to modify stem cell behaviour and differentiation. These channels are broadly expressed throughout human tissues and are involved in various cellular processes, such as the after-hyperpolarization in excitable cells, and also in differentiation processes. To this end, human induced pluripotent stem cells (hiPSCs) generated from plucked human hair keratinocytes have been exploited in vitro to recapitulate endoderm formation and, concomitantly, used to map the expression of the SK channel (SKCa) subtypes over time. Thus, we report the successful generation of definitive endoderm from hiPSCs of ectodermal origin using a highly reproducible and robust differentiation system. Furthermore, we provide the first evidence that SKCas subtypes are dynamically regulated in the transition from a pluripotent stem cell to a more lineage restricted, endodermal progeny.

L-Type Ca2+ Channels and SK Channels in Mouse Embryonic Stem Cells and Their Contribution to Cell Proliferation

“…These channels were blocked by the specific SK channel blocker apamin. The presence of SK channels in pluripotent stem cells is consistent with the fact that SK channels appear to be present in earlier stages of development (Illing et al 2013;Linta et al 2013).…”

Stepwise acquirement of hallmark neuropathology in FUS-ALS iPSC models depends on mutation type and neuronal aging

“…Vectors were co-transfected with packaging-defective helper plasmids into 293T cells using iPSCs from human hair keratinocytes were generated as described in (Illing et al, 2013;Linta et al, 2012;Stockmann and Lock, 2013) keratinocytes per well of a 6-well plate were infected with 5x10 7 proviral genome copies in EpiLife medium containing 8 mg/mL Polybrene (Sigma-Aldrich) at 2 sequent days. Another M A N U S C R I P T…”

Derivation and High Engraftment of Patient-Specific Cardiomyocyte Sheet Using Induced Pluripotent Stem Cells Generated From Adult Cardiac Fibroblast

“…If so, iPSC-derived cells may be more effective for treatment of myocardial injury if the iPSCs were created from cardiac-lineage cells, rather than (for example) dermal fibroblasts, which were the first cells used to generate iPSCs and continue to be the most common source of iPSCs for deriving cardiomyocytes (CMs). iPSCs have also been created from keratinocytes 11 , neuroprogenitor cells 12 , hemapoietic progenitor cells 13 , endothelial cells 14 , adipose stem cells 15 , peripheral blood cells 16 , hair follicle cells 17 , and cells obtained from the urine 18 . The experiments described in this report are the first to use iPSCs that have been engineered from cardiac fibroblasts.…”