In vitro cardiac safety evaluation for repurposed COVID-19 drugs by using human iPS cell-derived cardiomyocyte with gelatin fiber scaffold

Acquisition of pluripotency is driven largely at the transcriptional level by activators OCT4, SOX2, and NANOG that must in turn cooperate with diverse coactivators to execute stem cell-specific gene expression programs. Using a biochemically defined in vitro transcription system that mediates OCT4/SOX2 and coactivator-dependent transcription of the Nanog gene, we report the purification and identification of the dyskerin (DKC1) ribonucleoprotein complex as an OCT4/SOX2 coactivator whose activity appears to be modulated by a subset of associated small nucleolar RNAs (snoRNAs). The DKC1 complex occupies enhancers and regulates the expression of key pluripotency genes critical for self-renewal in embryonic stem (ES) cells. Depletion of DKC1 in fibroblasts significantly decreased the efficiency of induced pluripotent stem (iPS) cell generation. This study thus reveals an unanticipated transcriptional role of the DKC1 complex in stem cell maintenance and somatic cell reprogramming.

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Abstract 15596: Maturation of Hipsc-cm Electrophysiological and Contractile Function for Enhanced Sensitivity of Cardiac Safety Assays

Circulation ◽

10.1161/circ.142.suppl_3.15596 ◽

2020 ◽

Vol 142 (Suppl_3) ◽

Author(s):

Heather B Hayes ◽

Anthony M Nicolini ◽

Colin Arrowood ◽

Daniel Millard

Keyword(s):

Contractile Function ◽

Induced Pluripotent Stem Cell ◽

Force Frequency ◽

Cardiac Safety ◽

Electrophysiological Response ◽

Mechanical Conditioning ◽

Enhanced Sensitivity ◽

Frequency Relationship ◽

Positive Force

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have significantly advanced in vitro cardiac safety and disese modeling, yet remain an immature representation of human myocytes. Electrical or mechanical conditioning of hiPSC-CMs facilitates functional maturation, as measured by a positive force-frequency relationship, but current in vitro protocols require 2-4 weeks of conditioning. Using array-based contractility and local electrical stimulation, we detected functionally mature phenotypes and compound responses in hiPSC-CMs after only 48 hours of chronic pacing. To mature cardiomyocytes, hiPSC-CMs were cultured on 24- and 96-well MEA plates with a dedicated stimulation electrodes. Later, hiPSC-CMs were electrically or optically paced at 2Hz for 48 hours. Multimodal measures quantified contractile and electrophysiological responses to varied pacing rates and compound addition. After 48 hours of pacing, hiPSC-CMs displayed shortened repolarization timing compared to before chronic pacing (baseline: 423 +/- 21 ms; matured: 316 +/- 15 ms), without significant beat period changes (baseline: 1255 +/- 40 ms; matured: 1314 +/- 84 ms). Contractile beat amplitude was measured using array-based impedance during spontaneous beating and at increasing pacing rates (1, 1.2, 1.5, 2, and 2.5 Hz). Before chronic pacing, beat amplitude decreased with increasing pacing rate; after chronic pacing, the same wells displayed increased beat amplitudes with increasing pacing rate. The matured wells also showed enhanced sensitivity to positive inotropes, such as isoproterenol, digoxin, omecamtiv mecarbil, and dobutamine. Local extracellular action potentials (LEAP) further revealed altered electrophysiological response to ranolazine, a multichannel blocker. Unpaced control wells exhibited dose-dependent APD90 prolongation in response to ranolazine, whereas matured wells showed no APD90 change. Similar results were seen with 48 hour of optogenetic pacing at 2 Hz. Overall, hiPSC-CMs chronically paced for only 48 hours exhibited more mature functional phenotypes, including a positive force-frequnecy relationship, enhanced ionotrope sensitivity, and altered compound response.

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Anticancer activity in vitro and biological safety evaluation in vivo of Sika deer antler protein

Journal of Food Biochemistry ◽

10.1111/jfbc.12421 ◽

2017 ◽

Vol 41 (6) ◽

pp. e12421 ◽

Cited By ~ 2

Author(s):

Huihai Yang ◽

Lulu Wang ◽

Hang Sun ◽

Xiaofeng He ◽

Jing Zhang ◽

...

Keyword(s):

Anticancer Activity ◽

Sika Deer ◽

Safety Evaluation ◽

Deer Antler ◽

Biological Safety

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Using human iPS cell-derived enterocytes as novel in vitro model for the evaluation of human intestinal mucosal damage

Inflammation Research ◽

10.1007/s00011-018-1193-0 ◽

2018 ◽

Vol 67 (11-12) ◽

pp. 975-984 ◽

Cited By ~ 5

Author(s):

Satoshi Kondo ◽

Shota Mizuno ◽

Tadahiro Hashita ◽

Takahiro Iwao ◽

Tamihide Matsunaga

Keyword(s):

In Vitro Model ◽

Mucosal Damage ◽

Ips Cell ◽

Vitro Model

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Biological Safety Evaluation of 99mTc-DTPA-Ketoconazole for Diagnosis of Fungal Infection

Indonesian Journal of Physics and Nuclear Applications ◽

10.24246/ijpna.v2i1.1-8 ◽

2017 ◽

Vol 2 (1) ◽

pp. 1

Author(s):

Maula Eka Sriyani ◽

Hendris Wongso ◽

Eva Maria Widyasari ◽

Rizky Juwita Sugiharti ◽

Iim Halimah ◽

...

Keyword(s):

Single Dose ◽

Fungal Infection ◽

Radiochemical Purity ◽

Safety Evaluation ◽

Fungal Growth ◽

In Vitro Assays ◽

Toxic Response ◽

Biological Safety ◽

Safety Parameters

Infectious diseases have become one of the leading cause of mortality around the world, including in the Southeast Asia. One of the microbial that cause infection is fungi. Occasionally, deep-seated fungal infection is difficult to detect using conventional diagnosis methods and therefore leads to inaccurate detection. Our previous research was conducted in order to obtain the labeled compound of 99mTc-DTPA-Ketoconazole with a high radiochemical purity (98.40 ± 0.86%). Moreover, the in-vitro assays showed that 99mTc-DTPA-Ketoconazole can potentially bind to Candida albicans. On the other hand, in clinical routine use, diagnostic kit should be safe for the patients. Consequently, this research was conducted to determine the biological safety parameters of 99mTc-DTPA-Ketoconazole on the animal study, including single dose and acute toxicity test, sterility, and apirogenicity test. The results showed that both the single dose at 34.6 μCi and dose until 149 times of the single dose did not stimulate the toxic response to the animals. In addition, the sterility data revealed that there was no microbial growth after 7 days of incubation at 37°C as well as fungal growth after 14 days of incubation at 25°C. Furthermore, the apirogenicity test using rabbits revealed that there was no increase in temperature more than 0.6°C for each animal and not more than 1.5°C of total increase of temperature for all the animals. It is concluded that the 99mTc-DTPA-Ketoconazole is satisfy the requirements of biological safety of a radiopharmaceutical and therefore was acceptable for fungal detection in nuclear medicine.

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Development of an in vitro screening method for safety evaluation of nanomaterials

Bio-Medical Materials and Engineering ◽

10.3233/bme-2009-0559 ◽

2009 ◽

Vol 19 (1) ◽

pp. 19-27 ◽

Cited By ~ 3

Author(s):

Atsuko Matsuoka ◽

Agneta Önfelt ◽

Yoshie Matsuda ◽

Ryusuke Nakaoka ◽

Yuji Haishima ◽

...

Keyword(s):

Screening Method ◽

Safety Evaluation ◽

In Vitro Screening

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Albumin nanoparticles Preparation, Characterization and In-Vitro Safety Evaluation

Journal of Scientific Research in Science ◽

10.21608/jsrs.2021.69418.1054 ◽

2021 ◽

Vol 0 (0) ◽

pp. 0-0

Author(s):

dina youssef ◽

fatma sallam ◽

Taher Salaheldin ◽

Samah Darwish ◽

Abeer El-Metwally ◽

...

Keyword(s):

Safety Evaluation ◽

Albumin Nanoparticles

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Evaluation of low molecular weight polyethylenimine-introduced chitosan for gene delivery to mesenchymal stem cells

Materials Express ◽

10.1166/mex.2020.1780 ◽

2020 ◽

Vol 10 (7) ◽

pp. 1170-1176

Author(s):

Minchen Liu ◽

Yulan Hu ◽

Yi Feng

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Gene Delivery ◽

Transfection Efficiency ◽

A549 Cells ◽

Safety Evaluation ◽

Physicochemical Characteristics ◽

Hatching Rate

This study aimed to examine the transfection ability of polyethylenimine (PEI) (1800 Da)-grafted chitosan (10 kDa) (CP), a newly synthesized PEI derivative, in mesenchymal stem cells (MSCs). The safety evaluation of the complex/DNA was studied in vitro and in vivo. In addition, CP/pGL3 was applied to investigate the effects of transfection efficiency. In this study, CP/DNA can be formed with compatible physicochemical characteristics for gene delivery. CP cytotoxicity decreased in A549 cells. Moreover, a zebrafish embryo model was used for evaluating the safety in vivo. Compared to the PEI (25 kDa) group, the zebrafish hatching rate increased and the mortality rate decreased in the CP/DNA group, which provided an indication of the safety of CP. In comparison with chitosan (100 kDa)-PEI (1200 Da), CP's transfection efficiency was higher in both A549 cells and MSCs. This study aimed to lay the foundation for further applications of CP in gene delivery. Therefore, further gene therapy investigations of CP by using MSCs need to be performed.

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