Generation of patient-specific induced pluripotent stem cell lines from one patient with Jervell and Lange-Nielsen syndrome, one with type 1 long QT syndrome and two healthy relatives

Abstract

Four human iPSC cell lines (one Jervell and Lange-Nielsen Syndrome, one Long QT Syndrome-type 1 and two healthy controls) were generated from peripheral blood obtained from donors belonging to the same family. CytoTune™-iPS 2.0 Sendai Reprogramming Kit (containing OCT3/4, KLF4, SOX2 and cMYC as reprogramming factors) was used to generate all cell lines. The four iPSCs have normal karyotype, express pluripotency markers as determined by RT-PCR and flow cytometry and differentiated spontaneously in vitro into cells of the three germ layers, confirming their pluripotent capacity.

Extracellular Vesicles Derived from Induced Pluripotent Stem Cells Promote Renoprotection in Acute Kidney Injury Model

“…The experimental protocol was approved by the research ethics board of the Clementino Fraga Filho University Hospital (38583914.7.0000.5257/933.018, Rio de Janeiro, Brazil). Another iPSC line was obtained from peripheral blood, according to the protocol described in [16], and with the approval of the Research Ethics Committee of the National Institute of Cardiology (27044614.3.0000.5272, Rio de Janeiro, Brazil).…”

Long QT Syndrome Modelling with Cardiomyocytes Derived from Human-induced Pluripotent Stem Cells

“…[49,50] People with LQT1 and JLNS are affected by mutations in KCNQ1 that induce loss of functions in K v 7.1, an important potassium channel responsible for the slow delayed rectifier potassium current (I Ks ). Although I Ks is not straightforward to record in hiPSC-CMs due to their limited cell capacitance, which reduces the total current density, and to a late IKs onset during CM development,[51] partially mitigated by certain maturation strategies,[25] in vitro models of LQT1/JLNS were able to recapitulate the pathological phenotype and exhibited marked reductions in IKs magnitude when compared to hiPSC-CMs from healthy donors or to isogenic controls[52–54]; detailed investigation of I Ks with patch clamp also revealed the possibility for hiPSC-CMs to perfectly discriminate between homozygosity and heterozygosity. [51–55] The similarities between hiPSC-CMs and clinical disease features extend also to triggers of arrhythmias; as previously described in humans, an increased activation of the sympathetic cascade is a clear arrhythmogenic trigger for LQT1/JLNS.…”

Establishment of a heart-on-a-chip microdevice based on human iPS cells for the evaluation of human heart tissue function

“…HMDs may hold promise as a tool for "order-made" cardiac toxicity tests, since the HMDs can be prepared by patient-derived iPSCs requiring specific medications, such as anticancer drugs, for which the cardiac toxicity varies according to the biological backgrounds of the individuals. Furthermore, HMDs created with heart disease-specific human iPSCs can be used for the discovery of disease-specific treatment drugs through high-throughput screening of chemical compound libraries [37][38][39] . In addition to the validation of the basic system of the HMD, as shown in the present study, further medical applications of the HMDs as described above should be investigated in our next work, in which further optimization of the HMD system and experimental conditions such as the medium/buffer with defined chemical factors during drug tests, manipulations to firmly attach the 3D cardiac microtissue onto the microfluidic chip, and the identification of factors that might affect the tissue function and variation of data (e.g., cellular composition, 3D structure, and cellular alignments) should be conducted.…”