DISAGGREGATION OF PLANTS

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