DISAGGREGATION OF EXPLANTS

Flowchart nodes

• DISAGGREGATION OF EXPLANTS

• PRIMARY EXPLANT TECHNIQUE

• ENZYMATIC DISAGGREGATION

• MECHANICAL DISAGGREGATION

• DISSECTION

• FINE CHOPPING

• WASH BY SETTLING

• TRANSFER IN LAST WASH

• REMOVE BSS

• ADD THIN FILM OF MEDIUM

• INCUBATE FOR 2-3 WEEKS

• TISSUE SAMPLE

• CELL + TISSUE DEBRIS

• MORE CELLS +  TISSUE DEBRIS

• VIABLE CELLS + TISSUE DEBRIS

• VIABLE CELLS +  DAMAGED CELLS

• PRIMARY ADHERANT 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 CELLS

• EXPLANT OUTGROWTHS

• ENRICHED EPITHELIAL FRACTION

• DISPE CELL SUSPENSION CONTAINS ENRICHED FIBROBLASTSRSED

• PRIMARY CELL CULTURE

• PRIMARY CELL CULTURE