Culture and plate screening protocol for mammalian and insect cells

The plates represent a cross section of plates for different types of assays, cell growth and storage. We offer several formats: 96 and 24 wells to suit your needs.

Protocol for the screening of mammalian and insect cells – 96-well plates:

Material:
96-well plate, 2 ml, square wells, round bottom, individually wrapped with lid, sterile: 931134
96-well plate, 2 ml, square wells, pyramid bottom, individually wrapped with lid, sterile: 931133

Method:

  1. Maintain cell stocks in an appropriate growth medium. Divide cultures the day before transfection at an appropriate density to ensure growth of the log phase at the time of transfection.
  2. Add 500 µl / well of cell suspension. Optimal seed density depends on the cell line, please use the recommended density for the cell line used.
  3. Transfect the cells according to the established transfection protocol. Establish the ratio Transfection reagent / DNA / nutrients from what is used for the larger scale cultures.
  4. Seal the plates with plastic lids or gaskets and transfer them to a shaker overnight at 800 rpm and 37 ° C.
  5. Harvest cells at set time for larger scale cultures. Pellet the cells by centrifugation at 2,500 xg for 20 min at 4 ° C.
  6. Reserve culture medium or pellet depending on application and process downstream.

Notes:

The most critical factor in cell viability is aeration. Optimal results will be obtained using agitators with orbit diameters of 3 mm. We do not recommend working in 96-well plates using stirrers with standard diameters of 25 mm.
Thomson filter plates are an excellent add-on product for downstream purification applications.
96-well filter plate, 2 ml | 25µm Polypropylene: ref. 931919
Suggested maximum centrifugation: 3000 g.
Protocol for the screening of mammalian and insect cells – 24-well plates:

Material:
24-well plate, 10.4 mL, square wells, round bottom, individually wrapped with lid, sterile: 931568
24-well plate, 10.8 ml, square wells, pyramid bottom, individually wrapped with lid, sterile: 931571

Method:

  1. Maintain cell stocks in an appropriate growth medium. Divide cultures the day before transfection at an appropriate density to ensure growth of the log phase at the time of transfection.
  2. Place 4-5 ml / well of cell suspension. Optimal seed density depends on the cell line, please use the recommended density for the cell line used.
  3. Transfect the cells according to the established transfection protocol. Establish the ratio Transfection reagent / DNA / nutrients from what is used for the larger scale cultures.
  4. Seal the plates with plastic lids or gaskets and transfer them to a shaker overnight at 800 rpm and 37 ° C.
  5. Harvest cells at set time for larger scale cultures. Pellet the cells by centrifugation at 2,500 xg for 20 min at 4 ° C.
  6. Reserve culture medium or pellet depending on application and process downstream.

Notes:

The most critical factor in cell viability is aeration. Optimal results will be obtained using agitators with orbit diameters of 3 mm. We do not recommend working in 96-well plates using stirrers with standard diameters of 25 mm.
Thomson filter plates are an excellent add-on product for downstream purification applications.

Pat Lynch