DISAGGREGATION OF PLANTS

Nós do fluxograma

• DISAGGREGATION OF PLANTS

• PRIMARY AGGREGATION

• MECHANICAL DISAGGREGATION

• ENZYMATIC DISAGGREGATION

• ​​​​DISSECTION

• FINE CHOPPING  

• WASH BY SETTLING

• TRANSFER IN LAST WASH

• REMOVE BSS

• ADD THIN FILM OF MEDIUM

• INCUBATE FOR 2-3 WEEKS

• EXPLANT OUTGROWTHS

• TISSUE SAMPLE

• CELL TISSUE + DEBRIS

• MORE CELLS + TISSUE DEBRIS

• VIABLE CELLS + TISSUE DEBRIS

• VIABLE CELLS + DAMAGED CELLS

• PRIMARY ADHERENT CELL CULTURE

• BIOPSY IN MEDIUM + ANTIBIOTIC

• DISSECTION IN BSS  + ANTIBIOTIC

• CHOPPED TISSUE

• COMPLETE MEDIUM + COLLAGENASE

• DISAGGREGATED TISSUE + TISSUE RESIDUES

• DISPERSED CELLS + RESIDUAL CULTURE OF EPITHELIAL ELLS

• DISPERSED CELL SUSPENSION CONTAINS ENRICHED FIBROBLASTS

• PRIMARY CELL CULTURE

• ENRICHED EPITHELIAL FRACTION

• PRIMARY CELL CULTURE

• EDTA TREATMENT

• COLLAGENSE

• CENTRIFUGE

• RESUSPEND

• WASH

• PRIMARY CELL CULTURE