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Cardosins: A new and efficient plant enzymatic tool to dissociate neuronal cells for the establishment of cell cultures
Author(s) -
Duarte A.S.,
Rosa N.,
Duarte E.P.,
Pires E.,
Barros M.T.
Publication year - 2007
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.21259
Subject(s) - trypsin , cell culture , enzyme , embryonic stem cell , plant cell , depolarization , biochemistry , viability assay , biology , cell , microbiology and biotechnology , chemistry , biophysics , genetics , gene
In the present work, we examined the feasibility of using cardosins, plant aspartic‐proteinases from Cynara cardunculus L., to isolate cells from rat embryonic brain. Using morphological and functional assays, we compared cell cultures obtained with cardosins with those prepared with a well‐established trypsin protocol. Cardosins and trypsin dissociation produced cells with similar yield, viability, and GABA release in response to a depolarizing stimulus. However, cardosins‐dissociated cells appeared to recover faster in culture, as assessed by the MTT‐test and by the number and length of neurtites, suggesting that cardosins are less aggressive to neurons than trypsin. This feature might be helpful for research and medical purposes requiring fast manipulations of cells. Biotechnol. Bioeng. 2007;97: 991–996. © 2006 Wiley Periodicals, Inc.