Suppression of time-dependent decay by controlling the redox balance of human induced pluripotent stem cells suspended in a cryopreservation solution

Kagihiro, Masashi; Fukumori, Kazuhiro; Horiguchi, Ikki; Kim, Mee‐Hae; Kino‐oka, Masahiro

doi:10.1016/j.bej.2019.107465

Cited by 6 publications

(13 citation statements)

References 34 publications

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“…Figure 4 and Equation 6 define the following trend: The longer the filling time and the higher the required ratio of nonapoptotic cells are, the lower the probability of success becomes. This trend fits the experimental observations by Kagihiro et al (2020).…”

Section: Indicator Of Cell Survival and The Probability Of Filling supporting

confidence: 92%

“…Also, the consumption reaction of DMSO competes with the consumption reaction of H 2 O 2 , resulting in the increase of the ROS concentration. Kagihiro et al (2020) Kagihiro et al (2020) determined that the ROS concentration is a function of t S . In the experiment, the solution containing hiPS cells and DMSO as cryopreservation agent was filled into vials at t S = 0, 1, 2, 3, 4, 5, and 6 h. The measured/analyzed parameters included (i) apoptotic/non-apoptotic cells classified by flow cytometry with an active mitochondrial-specific dye (tetramethylrhodamine ethyl ester); (ii) ROS accumulation measured by flow cytometry with an oxidative stress detection dye (sample size for calculation in the order of 10 3 -10 4 counts); and (iii) the relative ROS accumulation level defined as the ratio of mean fluorescence intensity.…”

Section: Measurement Of Apoptosis In Hips Cellsmentioning

confidence: 99%

“…These experiments were performed five times each and at three temperature levels of 4 C, 25 C, and 37 C. At 25 C, two results were obtained with or without N-acetylcysteine (NAC) as an antioxidant. Kagihiro et al (2020) published the results at 4 C and 25 C with/without NAC. In the present study, all the results at 4 C, 25 C (with/without NAC), and 37 C were used.…”

Section: Measurement Of Apoptosis In Hips Cellsmentioning

confidence: 99%

“…In the present study, all the results at 4 C, 25 C (with/without NAC), and 37 C were used. See Kagihiro et al (2020) for the detail of the experiment. To evaluate the temporal change of the ROS ½ tS distribution from t S = 0, we adopted the indicator of Cohen's d (−), also known as the effect size (Cohen, 1988).…”

Section: Measurement Of Apoptosis In Hips Cellsmentioning

confidence: 99%

“…Using hiPS cells and mesenchymal stem cells, Kagihiro et al (2018) showed that the number of living cells decreased with filling time due to damages during the process. Kagihiro, Fukumori, Horiguchi, Kim, and Kino-oka (2020) further reported that the reactive oxygen species (ROS) accumulation induced apoptosis of cells in cryopreservation medium and caused the decrease of the non-apoptotic cell ratio during filling of hiPS cells. Thus, changes in cell quality need to be considered in determining the lot size.…”

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Apoptosis‐based method for determining lot sizes in the filling of human‐induced pluripotent stem cells

Sugiyama

Shiokaramatsu

Kagihiro

et al. 2020

J Tissue Eng Regen Med

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Standardization in process design and operation is needed in the commercial production of human-induced pluripotent stem (hiPS) cells. Lot sizing in the filling of hiPS cells into containers, a part of the preservation process, also needs to be standardized because of the temporal changes in cell quality during the process. Here, we present an apoptosis-based method that can determine lot sizes in the filling of hiPS cells considering temporal changes in cell quality. Two indicators were developed for (i) the cell quality change using reactive oxygen species (ROS) measurement and (ii) the cell survival and probability of filling success, which are parts of the lot-sizing problem. Using computational simulation, a map out of the optimal lot size was produced that minimized the expected production costs at a given cell demand and an acceptable change in cell quality. At a filling temperature of 4 C, the largest possible lot size was calculated as 6 L (corresponding to a filling time of 125 min). The results of a sensitivity analysis recommended cold filling or the addition of an antioxidant. The presented method is effective to determine the lot size considering the change in cell quality during filling. The study uniquely combines the experimental results with mathematical modeling and computational simulation techniques. The map out of the optimal lot size could guide the development of industrial filling processes of hiPS cells.

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Section: Indicator Of Cell Survival and The Probability Of Filling supporting

confidence: 92%

Section: Measurement Of Apoptosis In Hips Cellsmentioning

confidence: 99%

Section: Measurement Of Apoptosis In Hips Cellsmentioning

confidence: 99%

Section: Measurement Of Apoptosis In Hips Cellsmentioning

confidence: 99%

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confidence: 99%

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