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
N1 - Copyright:
Copyright 2013 Elsevier B.V., All rights reserved.
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.
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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 -