Cardosins: a new and efficient plant enzymatic tool to dissociate neuronal cells for the establishment of cell cultures.
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.
Duarte
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AS
CESAM and Departamento de Biologia, Universidade de Aveiro, Aveiro, Portugal.
Rosa
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N
Duarte
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EP
Pires
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E
Barros
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MT
D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000302:isolation & purification / Q000494:pharmacology*; D001921:Brain / Q000166:cytology* / Q000196:embryology; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D005240:Feasibility Studies; D005562:Formazans / Q000378:metabolism; D007150:Immunohistochemistry; D016501:Neurites / Q000502:physiology; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D010940:Plant Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D013778:Tetrazolium Salts / Q000378:metabolism; D013997:Time Factors; D014357:Trypsin / Q000494:pharmacology; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
D005562:Formazans; D010940:Plant Proteins; D013778:Tetrazolium Salts; C079782:MTT formazan; D005680:gamma-Aminobutyric Acid; D014357:Trypsin; D016282:Aspartic Acid Endopeptidases; C097455:aspartic proteinases, Cynara cardunculus