Generation of iPSC line iPSC-FH2.1 in hypoxic conditions from human foreskin fibroblasts

Questa, María; Romorini, Leonardo; Blüguermann, Carolina; Solari, Claudia; Neiman, Gabriel; Luzzani, Carlos; Scassa, María Élida; Sevlever, Gustavo; Guberman, Alejandra; Miriuka, Santiago Gabriel

doi:10.1016/j.scr.2015.12.026

Cited by 29 publications

(27 citation statements)

References 5 publications

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“…Cell lines, culture and differentiation hESCs WA09 (H9) were purchased from WiCell Research Institute (http://www.wicell.org) at low passages (p15 to p20). hiPSCs line FN2.1 has been previously derived from human foreskin fibroblasts at our laboratory in accordance with relevant guidelines and regulations and has been fully validated [64]. hPSCs lines were maintained on an inactivated mouse embryonic fibroblast (iMEF) feeder layer in medium comprised of Dulbecco's Modified Eagle's Medium/Ham's F12 (DMEM/F12, Gibco, http:// www.thermofisher.com/order/catalog/product/ 11330032) supplemented with 20% Knockout Serum Replacement (KSR, Gibco, http://www.thermofisher.…”

Section: Methodsmentioning

confidence: 99%

Regulation of cyclin E1 expression in human pluripotent stem cells and derived neural progeny

et al. 2018

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hPSCs: human pluripotent stem cells; hESCs: human embryonic stem cells; hiPSCs: human induced pluripotent stem cells; NP: neuroprogenitors; HF: human foreskin fibroblasts; MEFs: mouse embryonic fibroblasts; iMEFs: irradiated mouse embryonic fibroblasts; CDKs: cyclindependent kinases; CKIs: CDK inhibitors; CNS: central nervous system; Oct-4: Octamer-4; EB: embryoid body; AFP: Alpha-fetoprotein; cTnT: Cardiac Troponin T; MAP-2: microtubule-associated protein; TUJ-1: neuron-specific class III β-tubulin; bFGF: basic fibroblastic growth factor; PI3K: Phosphoinositide 3-kinase; KSR: knock out serum replacement; CM: iMEF conditioned medium; E8: Essential E8 medium.

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Section: Methodsmentioning

confidence: 99%

Regulation of cyclin E1 expression in human pluripotent stem cells and derived neural progeny

et al. 2018

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“…Induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were obtained by using the monolayer differentiation protocol described by Burridge et al 23 For this purpose, the FH2.1 induced pluripotent cells were used. 24 The assessment of the efficiency of cardiac differentiation was performed by using standard flow cytometry to determine the percentage of Troponin C + cells. The cells were maintained in Roswell Park Memorial Institute (RPMI) medium supplemented with B27 at 37°C and 5% CO2 until 35 days after the start of differentiation.…”

Section: Human Cardiomyocytes Derived From Induced Pluripotent Stem Cmentioning

confidence: 99%

Non-β-Blocking Carvedilol Analog, VK-II-86, Prevents Ouabain-Induced Cardiotoxicity

Gonano

Sepúlveda

Morell

et al. 2018

Circ J

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Background: It has been shown that carvedilol and its non β-blocking analog, VK-II-86, inhibit spontaneous Ca 2+ release from the sarcoplasmic reticulum (SR). The aim of this study is to determine whether carvedilol and VK-II-86 suppress ouabain-induced arrhythmogenic Ca 2+ waves and apoptosis in cardiac myocytes. Methods and Results: Rat cardiac myocytes were exposed to toxic doses of ouabain (50 µmol/L). Cell length (contraction) was monitored in electrically stimulated and non-stimulated conditions. Ouabain treatment increased contractility, frequency of spontaneous contractions and apoptosis compared to control cells. Carvedilol (1 µmol/L) or VK-II-86 (1 µmol/L) did not affect ouabain-induced inotropy, but significantly reduced the frequency of Ca 2+ waves, spontaneous contractions and cell death evoked by ouabain treatment. This antiarrhythmic effect was not associated with a reduction in Ca 2+ calmodulin-dependent protein kinase II (CaMKII) activity, phospholamban and ryanodine receptor phosphorylation or SR Ca 2+ load. Similar results could be replicated in human cardiomyocytes derived from stem cells and in a mathematical model of human myocytes. Conclusions: Carvedilol and VK-II-86 are effective to prevent ouabain-induced apoptosis and spontaneous contractions indicative of arrhythmogenic activity without affecting inotropy and demonstrated to be effective in human models, thus emerging as a therapeutic tool for the prevention of digitalis-induced arrhythmias and cardiac toxicity.

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“…The hiPSC cell line used in this study was generated in our lab from adult male fibroblasts and was previously described 22 . Cells were regularly cultured in 6 well plates in E8-Flex medium with Geltrex coating (all Thermo Fischer Scientific).…”

Section: D Cell Culturementioning

confidence: 99%

Single cell transfection of human induced pluripotent stem cells using a droplet-based microfluidic system

Perez

Sanluis-Verdes

Waisman

et al. 2020

Preprint

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Microfluidic tools have recently made possible many advances in biological and biomedical research. Research fields such as Physics, Engineering, Chemistry and Biology have combined to produce innovation in Microfluidics which has positively impacted on areas as diverse as nucleotide sequence, functional genomics, single-cell studies, single molecules assays, and biomedical diagnostics. Among these areas regenerative medicine and stem cells have benefited from Microfluidics due to these tools have had a profound impact on their applications. In the study, we present a high-performance droplet-based system for transfecting individual humaninduced pluripotent stem cells. We show that this system has great efficiency in single cells and captured droplets, similar to other microfluidic methods and lower cost. We demonstrate that this microfluidic approach can be associated with the PiggyBac transposase-based system to increase its transfection efficiency. Our results provide a starting point for subsequent applications in more complex transfection systems, single-cell differentiation interactions, cell subpopulations, cell therapy, among other potential applications.

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Generation of iPSC line iPSC-FH2.1 in hypoxic conditions from human foreskin fibroblasts

Cited by 29 publications

References 5 publications

Regulation of cyclin E1 expression in human pluripotent stem cells and derived neural progeny

Regulation of cyclin E1 expression in human pluripotent stem cells and derived neural progeny

Non-β-Blocking Carvedilol Analog, VK-II-86, Prevents Ouabain-Induced Cardiotoxicity

Single cell transfection of human induced pluripotent stem cells using a droplet-based microfluidic system

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