Unprecedented cell-selection using ultra-quick freezing combined with aquaporin expression

Yasuhiro Kato, Takayuki Miyauchi, Yoichiro Abe, Dušan Kojić, Manami Tanaka, Nana Chikazawa, Yuki Nakatake, Shigeru Ko, Daisuke Kobayashi, Akihiro Hazama, Shoko Fujiwara, Tatsuya Uchida, Masato Yasui

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Freezing is usually used for preservation and storage of biological samples; however, this process may have some adverse effects such as cell membrane damage. Aquaporin (AQP), a water channel protein, has been suggested to play some roles for cryopreservation although its molecular mechanism remains unclear. Here we show that membrane damage caused by ultra-quick freezing is rescued by the expression of AQP4. We next examine if the expression of AQP combined with ultra-quick freezing can be used to select cells efficiently under freezing conditions where most cells are died. CHO cells stably expressing AQP4 were exclusively selected from mixed cell cultures. Having identified the increased expression of AQP4 during ES cell differentiation into neuro-ectoderm using bioinformatics, we confirmed the improved survival of differentiated ES cells with AQP4 expression. Finally we show that CHO cells transiently transfected with Endothelin receptor A and Aqp4 were also selected and concentrated by multiple cycles of freezing/thawing, which was confirmed with calcium imaging in response to endothelin. Furthermore, we found that the expression of AQP enables a reduction in the amount of cryoprotectants for freezing, thereby decreasing osmotic stress and cellular toxicity. Taken together, we propose that this simple but efficient and safe method may be applicable to the selection of mammalian cells for applications in regenerative medicine as well as cell-based functional assays or drug screening protocols.

Original languageEnglish
Article numbere87644
JournalPLoS One
Volume9
Issue number2
DOIs
Publication statusPublished - 2014 Feb 18

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Aquaporins
aquaporins
Freezing
freezing
CHO Cells
cells
endothelins
Biological Preservation
Endothelin A Receptors
Ectoderm
Preclinical Drug Evaluations
Thawing
Regenerative Medicine
Cryopreservation
Endothelins
Osmotic Pressure
Cell membranes
Bioinformatics
Computational Biology
Cell culture

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Unprecedented cell-selection using ultra-quick freezing combined with aquaporin expression. / Kato, Yasuhiro; Miyauchi, Takayuki; Abe, Yoichiro; Kojić, Dušan; Tanaka, Manami; Chikazawa, Nana; Nakatake, Yuki; Ko, Shigeru; Kobayashi, Daisuke; Hazama, Akihiro; Fujiwara, Shoko; Uchida, Tatsuya; Yasui, Masato.

In: PLoS One, Vol. 9, No. 2, e87644, 18.02.2014.

Research output: Contribution to journalArticle

Kato, Y, Miyauchi, T, Abe, Y, Kojić, D, Tanaka, M, Chikazawa, N, Nakatake, Y, Ko, S, Kobayashi, D, Hazama, A, Fujiwara, S, Uchida, T & Yasui, M 2014, 'Unprecedented cell-selection using ultra-quick freezing combined with aquaporin expression', PLoS One, vol. 9, no. 2, e87644. https://doi.org/10.1371/journal.pone.0087644
Kato, Yasuhiro ; Miyauchi, Takayuki ; Abe, Yoichiro ; Kojić, Dušan ; Tanaka, Manami ; Chikazawa, Nana ; Nakatake, Yuki ; Ko, Shigeru ; Kobayashi, Daisuke ; Hazama, Akihiro ; Fujiwara, Shoko ; Uchida, Tatsuya ; Yasui, Masato. / Unprecedented cell-selection using ultra-quick freezing combined with aquaporin expression. In: PLoS One. 2014 ; Vol. 9, No. 2.
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