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

4 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

Access to Document

10.1371/journal.pone.0042485

Cite this

@article{da84993293e44e05b17fcf155d26c5de,

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

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.",

author = "Hideyuki Terazono and Hyonchol Kim and Masahito Hayashi and Akihiro Hattori and Fumimasa Nomura and Tomoyuki Kaneko and Kenji Yasuda",

year = "2012",

month = aug,

day = "3",

doi = "10.1371/journal.pone.0042485",

language = "English",

volume = "7",

journal = "PLoS One",

issn = "1932-6203",

publisher = "Public Library of Science",

number = "8",

}

TY - JOUR

T1 - A non-destructive culturing and cell sorting method for cardiomyocytes and neurons using a double alginate layer

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

N2 - 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.

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.

UR - http://www.scopus.com/inward/record.url?scp=84864535440&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84864535440&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0042485

DO - 10.1371/journal.pone.0042485

M3 - Article

C2 - 22870332

AN - SCOPUS:84864535440

VL - 7

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 8

M1 - e42485

ER -

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

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this