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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.

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.

Materials
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