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iCell GABANeurons KCNT1 (P924L), 01434

Kit Size

Catalog #: R1166

Cells Only

Catalog #: C1173
Catalog #: C1173

Epilepsy differentiated from human iPS cells, frozen

From
$2,545.00
Catalog # P924L-01434
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Product Overview

Mutations in KCNT1, encoding the sodium-activated potassium channel (Slack, KNa1.1), are associated with infant and late-onset focal epileptic encephalopathies, including malignant migrating partial seizures of infancy (MMPSI) and autosomal-dominant nocturnal frontal lobe epilepsy (ADNFLE). Discovery and validation of therapeutics that target KCNT1-related epilepsy disorders can greatly benefit from relevant and accessible human cellular models.

iCell GABANeurons KCNT1 (P924L) feature a homozygous P924L mutation in the KNa1.1 receptor resulting in the gain of function increase in outward potassium channel current, a known phenotype associated with frontal lobe epilepsy. The KCNT1 P924L mutation was engineered into induced pluripotent stem cells, followed by differentiation into cortical neurons that exhibit a primarily GABAergic phenotype.

Features of iCell GABANeurons KCNT1 (P924L):

  • >95% pure neuronal population of primarily GABAergic neurons
  • Increased KNa1.1 potassium current
  • Hyperactive neural networks that can be suppressed by Quinidine
  • Enhanced phenotype when co-cultured with iCell GlutaNeurons
  • Consistent lot-to-lot GABA phenotypes and cell viability
  • Isogenic iCell GABANeurons, 01434 are available

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Our specialists are here to help you find the best product for your application.

Call Us

(877) 320-6688

 

OR

Our regular business hours are 9:00am to 5:00pm Central Time (USA)

Performance Data

KCNT1 (P924L) iCell GABANeurons Viability is Comparable Isogenic Control.

Isogenic control (iCell GABANeurons, 01434) and KCNT1 (P924L) iCell GABANeurons were cultured at equivalent cell densities for 7 days and analyzed for cell viability. A) Representative images of KCNT1 (P924L) GABANeurons and isogenic control GABANeurons stained with Calcein AM (green) and DAPI (blue). B) Quantification of Calcein AM showed similar numbers of viable KCNT1(P924L) iCell GABANeurons compared to isogenic control.

Figure 1: KCNT1 (P924L) iCell GABANeurons Viability is Comparable Isogenic Control

Figure 2: Increased Neurite Growth and Branching for KCNT1 (P924L) iCell GABANeurons

Increased Neurite Growth and Branching for KCNT1 (P924L) iCell GABANeurons.

Isogenic control (iCell GABANeurons, 01434) and iCell GABANeurons KCNT1 (P924L) were cultured at equivalent cell densities and analyzed for neurite outgrowth by high content imaging. A) Representative brightfield images of KCNT1 (P924L) iCell GABANeurons and isogenic control GABANeurons at 7 days in culture. B) KCNT1(P924L) iCell GABANeurons display increased mean neurite outgrowth, branch number and maximum process length compared to isogenic control.

iCell GABANeurons KCNT1 (P924L) Display Expected Action Potential Waveform Pathophysiology.

A) Representative whole-cell current-clamp patch recordings of evoked action potentials (AP) at multiple currents (inset) for iCell GABANeurons KCNT1 (P924L) and isogenic control (iCell GABANeurons, 01434). Whole-cell current-clamp recordings show that (B) AP spike-width is decreased, (C) fast after-hyperpolarization (AHP) is increased, and (D) total evoked APs via current steps (upper graph insets) are increased in KCNT1 (P924L) iCell GABANeurons compared to isogenic GABANeurons. These pathological characteristics align with the ‘gain-of-function’ phenotype observed in many KCNT1 mutations, resulting in a hyper-excitable cell-state.

Figure 3: iCell GABANeurons KCNT1 (P924L) Display Expected Action Potential Waveform Pathophysiology

Figure 4: Network Activity is Increased in iCell GABANeurons KCNT1 (P924L)

Network Activity is Increased in iCell GABANeurons KCNT1 (P924L).

KCNT1 (P924L) GABANeurons and isogenic control (iCell GABANeurons) were cultured and monitored for network activity via MEA. A) Representative raster plots of KCNT1 (P924L) and isogenic control neurons. Analysis of firing rate and intensity show that KCNT1 (P924L) GABANeurons have increased synchrony as indicated by burst intensity (B) and an increased mean firing rate (C) compared to isogenic GABANeurons. These data suggest KCNT1 (P924L) neurons are more “epileptic” than isogenic neurons.

Quinidine Attenuates Activity in KCNT1 (P924L) iCell GABANeurons.

iCell GABANeurons KCNT1 (P924L) or isogenic control iCell GABANeurons were cultured for 15 days prior to treatment with Quinidine (50, 100, or 200 µM), a KCNT1 channel blocker. Following 24 hours, neural activity was recorded by multielectrode array (MEA) and analyzed for mean firing rate (Hz) and firing synchrony. Quinidine resulted in a dose-dependent reduction in mean firing rate and synchrony of KCNT1 (P924L) GABANeuron cultures.

Figure 5: Quinidine Attenuates Activity in KCNT1 (P924L) iCell GABANeurons

Figure 6: Co-culture with iCell GlutaNeurons Enhances KCNT1 (P924L) GABANeuron Phenotype

Co-culture with iCell GlutaNeurons Enhances KCNT1 (P924L) GABANeuron Phenotype.

iCell GlutaNeurons were co-cultured with either iCell GABANeurons KCNT1 (P924L) or isogenic control iCell GABANeurons at varying ratios of GlutaNeuron:GabaNeuron. Neural activity was monitored via multielectrode array (MEA) following 19 days in culture. Activity traces show that increasing the GABANeuron ratio results in increased neural network burst frequency compared to GlutaNeurons alone. Under all co-culture ratios, KCNT1 (P942L) GABANeurons showed increased network burst frequency compared to cultures containing isogenic GABANeurons.

Enhanced Disease Phenotype in iCell GABANeurons KCNT1 (P924L) when Co-Cultured with iCell GlutaNeurons.

iCell GlutaNeurons were co-cultured with either iCell GABANeurons KCNT1 (P924A) or isogenic control iCell GABANeurons at a ratio of 1:3. Neural activity was monitored via multielectrode array (MEA) following 19 days in culture. A) Raster plots show network burst frequency is increased in KCNT1 (P924L) cultures compared to isogenic control GABANeurons or iCell GlutaNeurons in monoculture.

Figure 7: Enhanced Disease Phenotype in iCell GABANeurons KCNT1 (P924L) when Co-Cultured with iCell GlutaNeurons

Product Highlights

Applications:

  • Drug target and therapeutic screening
  • Epilepsy disease modeling
  • Seizurogenic compound testing
  • Autosomal-dominant nocturnal frontal lobe epilepsy
  • Malignant migrating partial seizures of infancy