Coating of tissue culture dishes: Use 3 to 10 microgram per 1 cm2 dish area. For each cell type and plate type an optimization may be required. Dilute in 10 mM HCl or PBS. Apply the diluted collagen to a tissue culture plate and allow to dry overnight in a laminar flow hood with the plate lid open. Aspirate gently the remaining liquid from the coated wells. Wash twice gently with PBS.
To prepare a collagen gel the fibrillogenesis buffer contains 162 mM sodium phosphate dibasic (Na2HPO4) adjusted to pH 11.2 with 10 N NaOH, filter sterilized. Mix 9 volumes of Collagen at a concentration of 0.3%-1% with 1 volume of fibrillogenesis buffer. Mix well and incubate for 4 to 16 hours at 25°C to 27 °C. Higher concentration of collagen solution might need optimization. Thiis preparation will not spontaneously form a gel in PBS or cell culture medium.
Preparation of collagen sponge: concentrate gel to 10 mg/ml - 100 mg/ml by centrifugation, freeze dry. Crosslink by common methods found in the scientific literature such as 1-Ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC), Glutaraldehyde or thermal dehydration.
Other forms: membranes, sheets, fibers can be prepared by methods commonly used with other types of collagen.