Cardosins: A new and efficient plant enzymatic tool to dissociate neuronal cells for the establishment of cell cultures

• Published in: Biotechnology and bioengineering Vol. 97; no. 4; pp. 991 - 996
• Main Authors: Duarte, A.S., Rosa, N., Duarte, E.P., Pires, E., Barros, M.T.
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.07.2007; Wiley; Wiley Subscription Services, Inc
• Subjects: Animals; Aspartic Acid Endopeptidases - isolation & purification; Aspartic Acid Endopeptidases - pharmacology; Biological and medical sciences; Biotechnology; Brain - cytology; Brain - embryology; cardosins; Cell culture; Cell Culture Techniques; Cell Survival - drug effects; cell viability; Cells, Cultured; Comparative analysis; Cynara cardunculus; Enzymes; Feasibility Studies; Formazans - metabolism; Fundamental and applied biological sciences. Psychology; gamma-Aminobutyric Acid - secretion; Immunohistochemistry; Neurites - physiology; neuronal cultures; Neurons; Neurons - cytology; Neurons - metabolism; Plant Proteins - isolation & purification; Plant Proteins - pharmacology; Rats; Rats, Wistar; Tetrazolium Salts - metabolism; Time Factors; tissue dissociation; trypsin; Trypsin - pharmacology; Peptidases; Cell culture; Tissue; Trypsin; cell viability; Serine endopeptidases; Enzyme; Hydrolases; neuronal cultures; tissue dissociation; Viability; cardosins
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.21259

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.