Enhanced single-cell viability using 30Kc6 for efficient expansion of human induced pluripotent stem cells

Jina Ryu; Sang Wook Park; Hee Ho Park; Tai Hyun Park

doi:10.1016/j.procbio.2019.01.017

Enhanced single-cell viability using 30Kc6 for efficient expansion of human induced pluripotent stem cells

Jina Ryu, Sang Wook Park, Hee Ho Park, Tai Hyun Park

Department of Nutritional Science & Food Management

Research output: Contribution to journal › Article › peer-review

Abstract

Human pluripotent stem cells (hPSCs), such as human induced pluripotent stem cells (hiPSCs) and human embryonic stem cells (hESCs), have been widely used in stem cell research. hPSCs form colonies in culture dishes and are passaged as clumps. If the cells are dissociated into single cells, epithelial structures are disrupted, and rapid apoptosis is induced. The low viability of single cells restricts the passaging and expansion of hPSCs. Here, in order to enhance the single cell viability of hPSCs, we adopted an apoptotic strategy using the 30 Kc6 gene which inhibits mitochondrial apoptosis. With the expression of the 30 Kc6 gene in hiPSCs, no issues related to pluripotency or differentiation occurred. The hiPSC-30 Kc6 showed higher viability after the induction of apoptosis compared with control hiPSCs. Furthermore, hiPSC-30 Kc6 formed more colonies when the cells were enzymatically dissociated into single cells. Taken together, this study demonstrates that the 30 Kc6 gene could facilitate the passage and expansion of hiPSCs, potentially preventing the dissociation-induced apoptosis of single hPSCs.

Original language	English
Pages (from-to)	161-168
Number of pages	8
Journal	Process Biochemistry
Volume	78
DOIs	https://doi.org/10.1016/j.procbio.2019.01.017
State	Published - Mar 2019

Bibliographical note

Funding Information:
This study was supported by the National Research Foundation of Korea (NRF) funded by the Korean government (MSIT) (2017M3A9C6031786, 2017M3A9C6031798). This study was supported by 2018 Research Grant from Kangwon National University. This resesarch was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2018R1D1A1B07050422).

Funding Information:
This study was supported by the National Research Foundation of Korea (NRF) funded by the Korean government (MSIT) ( 2017M3A9C6031786 , 2017M3A9C6031798 ). This study was supported by 2018 Research Grant from Kangwon National University . This resesarch was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education ( NRF-2018R1D1A1B07050422 ).

Publisher Copyright:
© 2019 Elsevier Ltd

Keywords

Enzymatic dissociation
Single cell viability
hiPSC
hiPSC-30Kc6

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10.1016/j.procbio.2019.01.017

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title = "Enhanced single-cell viability using 30Kc6 for efficient expansion of human induced pluripotent stem cells",

abstract = "Human pluripotent stem cells (hPSCs), such as human induced pluripotent stem cells (hiPSCs) and human embryonic stem cells (hESCs), have been widely used in stem cell research. hPSCs form colonies in culture dishes and are passaged as clumps. If the cells are dissociated into single cells, epithelial structures are disrupted, and rapid apoptosis is induced. The low viability of single cells restricts the passaging and expansion of hPSCs. Here, in order to enhance the single cell viability of hPSCs, we adopted an apoptotic strategy using the 30 Kc6 gene which inhibits mitochondrial apoptosis. With the expression of the 30 Kc6 gene in hiPSCs, no issues related to pluripotency or differentiation occurred. The hiPSC-30 Kc6 showed higher viability after the induction of apoptosis compared with control hiPSCs. Furthermore, hiPSC-30 Kc6 formed more colonies when the cells were enzymatically dissociated into single cells. Taken together, this study demonstrates that the 30 Kc6 gene could facilitate the passage and expansion of hiPSCs, potentially preventing the dissociation-induced apoptosis of single hPSCs.",

keywords = "Enzymatic dissociation, Single cell viability, hiPSC, hiPSC-30Kc6",

author = "Jina Ryu and Park, {Sang Wook} and Park, {Hee Ho} and Park, {Tai Hyun}",

note = "Funding Information: This study was supported by the National Research Foundation of Korea (NRF) funded by the Korean government (MSIT) (2017M3A9C6031786, 2017M3A9C6031798). This study was supported by 2018 Research Grant from Kangwon National University. This resesarch was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2018R1D1A1B07050422). Funding Information: This study was supported by the National Research Foundation of Korea (NRF) funded by the Korean government (MSIT) ( 2017M3A9C6031786 , 2017M3A9C6031798 ). This study was supported by 2018 Research Grant from Kangwon National University . This resesarch was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education ( NRF-2018R1D1A1B07050422 ). Publisher Copyright: {\textcopyright} 2019 Elsevier Ltd",

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doi = "10.1016/j.procbio.2019.01.017",

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AU - Park, Sang Wook

AU - Park, Hee Ho

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N1 - Funding Information: This study was supported by the National Research Foundation of Korea (NRF) funded by the Korean government (MSIT) (2017M3A9C6031786, 2017M3A9C6031798). This study was supported by 2018 Research Grant from Kangwon National University. This resesarch was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2018R1D1A1B07050422). Funding Information: This study was supported by the National Research Foundation of Korea (NRF) funded by the Korean government (MSIT) ( 2017M3A9C6031786 , 2017M3A9C6031798 ). This study was supported by 2018 Research Grant from Kangwon National University . This resesarch was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education ( NRF-2018R1D1A1B07050422 ). Publisher Copyright: © 2019 Elsevier Ltd

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N2 - Human pluripotent stem cells (hPSCs), such as human induced pluripotent stem cells (hiPSCs) and human embryonic stem cells (hESCs), have been widely used in stem cell research. hPSCs form colonies in culture dishes and are passaged as clumps. If the cells are dissociated into single cells, epithelial structures are disrupted, and rapid apoptosis is induced. The low viability of single cells restricts the passaging and expansion of hPSCs. Here, in order to enhance the single cell viability of hPSCs, we adopted an apoptotic strategy using the 30 Kc6 gene which inhibits mitochondrial apoptosis. With the expression of the 30 Kc6 gene in hiPSCs, no issues related to pluripotency or differentiation occurred. The hiPSC-30 Kc6 showed higher viability after the induction of apoptosis compared with control hiPSCs. Furthermore, hiPSC-30 Kc6 formed more colonies when the cells were enzymatically dissociated into single cells. Taken together, this study demonstrates that the 30 Kc6 gene could facilitate the passage and expansion of hiPSCs, potentially preventing the dissociation-induced apoptosis of single hPSCs.

AB - Human pluripotent stem cells (hPSCs), such as human induced pluripotent stem cells (hiPSCs) and human embryonic stem cells (hESCs), have been widely used in stem cell research. hPSCs form colonies in culture dishes and are passaged as clumps. If the cells are dissociated into single cells, epithelial structures are disrupted, and rapid apoptosis is induced. The low viability of single cells restricts the passaging and expansion of hPSCs. Here, in order to enhance the single cell viability of hPSCs, we adopted an apoptotic strategy using the 30 Kc6 gene which inhibits mitochondrial apoptosis. With the expression of the 30 Kc6 gene in hiPSCs, no issues related to pluripotency or differentiation occurred. The hiPSC-30 Kc6 showed higher viability after the induction of apoptosis compared with control hiPSCs. Furthermore, hiPSC-30 Kc6 formed more colonies when the cells were enzymatically dissociated into single cells. Taken together, this study demonstrates that the 30 Kc6 gene could facilitate the passage and expansion of hiPSCs, potentially preventing the dissociation-induced apoptosis of single hPSCs.

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Enhanced single-cell viability using 30Kc6 for efficient expansion of human induced pluripotent stem cells

Abstract

Bibliographical note

Keywords

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