Critical Functionality Effects from Storage Temperature on Human Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium Cell Suspensions

Scientific Reports
Shohei KitahataMasayo Takahashi

Abstract

Human induced pluripotent stem cell (hiPSC)-derived retinal pigment epithelium (hiPSC-RPE) cells suspension have the potential for regenerative treatment. However, practical regenerative applications with hiPSC-RPE cells require the development of simple and cost-effective non-freezing preservation methods. We investigated the effect of non-freezing temperatures on suspended hiPSC-RPE cells in various conditions and analysed mechanisms of cell death, anoikis, Rho GTPases, hypoxia, microtubule destruction, and cell metabolism. Cells stored at 37 °C had the lowest viability due to hypoxia from high cell metabolism and cell deposits, and cells preserved at 4 °C were damaged via microtubule fragility. Cell suspensions at 16 °C were optimal with drastically reduced apoptosis and negligible necrosis. Moreover, surviving cells proliferated and secreted key proteins normally, compared to cells without preservation. hiPSC-RPE cell suspensions were optimally preserved at 16 °C. Temperatures above or below the optimal temperature decreased cell viability significantly yet differentially by mechanisms of cell death, cellular metabolism, microtubule destruction, and oxygen tension, all relevant to cell conditions. Surviving cells are expect...Continue Reading

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Citations

Apr 2, 2019·Investigative Ophthalmology & Visual Science·Kenichi MakabeMasayo Takahashi
Nov 7, 2019·Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan·Hiroshi Ueda
Dec 16, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Rakibul IslamTor Paaske Utheim
May 5, 2021·Nature Methods·Rajarshi Pal
Sep 7, 2021·Frontiers in Medicine·Ayyad Zartasht KhanJon Roger Eidet
Dec 3, 2021·Biomacromolecules·Zhengyu PiaoByeongmoon Jeong

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Methods Mentioned

BETA
flow cytometry
ELISA
enzyme-linked immunosorbent assay
GTPases
fluorescence-activated cell sorting
Assay

Software Mentioned

ImageJ
SPSS

Related Concepts

Related Feeds

Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis

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