Gelatin Coating Protocol for Cell Culture

Gelatin is a molecular derivative of collagen and is synthesized from the alkaline digestion of collagen from bovine skin. Gelatin is a hydrocolloid and is rich in glycine, proline and hydroxyproline, which imparts structural stability. Gelatin offers an ideal cell-attachment substrate for cell culture. The optimal concentration depends on cell type and research objective.

Important Notes before Gelatin Coating

• Ensure cells are healthy and in an adequate number.
• Gelatin solutions need to completely liquefy at 37 °C before coating.
• Gelatin solutions may be stored at 4 - 8 °C indefinitely; elevated temperatures cause hydrolysis and loss of integrity.
• The typical surface coverage concentration is 100 – 200 µg/cm². Surface coverage concentration may differ with the cells being cultured. Optimal conditions for attachment must be determined for each cell line and application.
• Do not over-dry post coating as this disrupts the structure of gelatin and affects cell attachment.

Gelatin coating protocol for Cell culture

1. Prepare a 2% (w/v) solution by dissolving gelatin in tissue-culture grade water.
2. Sterilize by autoclaving at 121 °C, 15 psi for 30 minutes.
3. Coat culture surface with 5-10 µL gelatin solution/cm² (i.e., 100-200 µg/cm²).
4. Allow to dry at least 2 hours before introducing cells and medium.
5. Alternatively, a gelatin solution (Product No. G1393) can be used to directly coat the cultureware.

Table 1. Gelatins for cell attachment.

Product No	Description	Coating concentration	Dilute in
G1393	Gelatin solution	100‑200 μg/cm2	Sterile water
G9136	Gelatin from porcine skin	100‑200 μg/cm2	Warm basal medium (without serum)
G1890	Gelatin from porcine skin	100‑200 μg/cm2	Warm water
G7041	Gelatin from cold water fish skin	100‑200 μg/cm2
G9391	Gelatin from bovine skin	100‑200 μg/cm2

Frequently Asked Questions for Coating Culture ware with gelatin

• What is the purity of gelatin products?
  We do not determine the purity of fibronectin solutions. All our fibronectin powders are 70 - 90% pure.
  
  
• What is the best solvent to dilute gelatin powders?
  The solvents that are compatible to dilute gelatins are provided in Table 1.
  
  
• How long can gelatin be stored?
  Gelatin solutions can be stored indefinitely at 2 - 8 °C. Gelatin-coated plates can be stored at 2 - 8 °C for up to 4 weeks provided they are sealed well to prevent contamination and/or over-drying up of gelatin. Do not use the product if discoloration or cracks appear on the surface of the coated material.
  
  
• What is the significance of Bloom number for gelatins?
  Bloom number was introduced by Oscar T. Bloom and indicates the stiffness of a gelatin film. It was first measured using a Bloom Gelometer by a controlled process to measure the rigidity of a gelatin film. A higher number indicates a stiffer product. The Bloom number/Bloom strength helps in choosing the right gelatin product for your applications.