A non-destructive culturing and cell sorting method for cardiomyocytes and neurons using a double alginate layer

Hideyuki Terazono; Hyonchol Kim; Masahito Hayashi; Akihiro Hattori; Fumimasa Nomura; Tomoyuki Kaneko; Kenji Yasuda

doi:10.1371/journal.pone.0042485

A non-destructive culturing and cell sorting method for cardiomyocytes and neurons using a double alginate layer

Hideyuki Terazono, Hyonchol Kim, Masahito Hayashi, Akihiro Hattori, Fumimasa Nomura, Tomoyuki Kaneko, Kenji Yasuda^*

^*Corresponding author for this work

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

6 Citations (Scopus)

Abstract

A non-destructive method of collecting cultured cells after identifying their in situ functional characteristics is proposed. In this method, cells are cultivated on an alginate layer in a culture dish and released by spot application of a calcium chelate buffer that locally melts the alginate layer and enables the collection of cultured cells at the single-cell level. Primary hippocampal neurons, beating human embryonic stem (hES) cell-derived cardiomyocytes, and beating hES cell-derived cardiomyocyte clusters cultivated on an alginate layer were successfully released and collected with a micropipette. The collected cells were recultured while maintaining their physiological function, including beating, and elongated neurites. These results suggest that the proposed method may eventually facilitate the transplantation of ES- or iPS-derived cardiomyocytes and neurons differentiated in culture.

Original language	English
Article number	e42485
Journal	PloS one
Volume	7
Issue number	8
DOIs	https://doi.org/10.1371/journal.pone.0042485
Publication status	Published - 2012 Aug 3
Externally published	Yes

ASJC Scopus subject areas

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

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abstract = "A non-destructive method of collecting cultured cells after identifying their in situ functional characteristics is proposed. In this method, cells are cultivated on an alginate layer in a culture dish and released by spot application of a calcium chelate buffer that locally melts the alginate layer and enables the collection of cultured cells at the single-cell level. Primary hippocampal neurons, beating human embryonic stem (hES) cell-derived cardiomyocytes, and beating hES cell-derived cardiomyocyte clusters cultivated on an alginate layer were successfully released and collected with a micropipette. The collected cells were recultured while maintaining their physiological function, including beating, and elongated neurites. These results suggest that the proposed method may eventually facilitate the transplantation of ES- or iPS-derived cardiomyocytes and neurons differentiated in culture.",

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AU - Terazono, Hideyuki

AU - Kim, Hyonchol

AU - Hayashi, Masahito

AU - Hattori, Akihiro

AU - Nomura, Fumimasa

AU - Kaneko, Tomoyuki

AU - Yasuda, Kenji

PY - 2012/8/3

Y1 - 2012/8/3

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AB - A non-destructive method of collecting cultured cells after identifying their in situ functional characteristics is proposed. In this method, cells are cultivated on an alginate layer in a culture dish and released by spot application of a calcium chelate buffer that locally melts the alginate layer and enables the collection of cultured cells at the single-cell level. Primary hippocampal neurons, beating human embryonic stem (hES) cell-derived cardiomyocytes, and beating hES cell-derived cardiomyocyte clusters cultivated on an alginate layer were successfully released and collected with a micropipette. The collected cells were recultured while maintaining their physiological function, including beating, and elongated neurites. These results suggest that the proposed method may eventually facilitate the transplantation of ES- or iPS-derived cardiomyocytes and neurons differentiated in culture.

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A non-destructive culturing and cell sorting method for cardiomyocytes and neurons using a double alginate layer

Abstract

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