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MEA User Manual
5.4.9 Coating with Collagen
Coating with collagen is useful for short-term cultures. It tends to detach from the surface if
used for long-term cultures.
Materials
•
DMEM Dulbecco’s Modified Eagle Media (DMEM) / F12
•
0.01 N Hydrochloric acid, pH 3.0
•
Acid-soluble type I collagen solution(3mg/ml, pH3.0) Cellmatrix Type I-A (Nitta Gelatin Inc.)
(Gibco/Invitrogen, 21331-020)
Preparation buffer
•
200 mM HEPES in 0.08 N NaOH
Collagen solution
1. Add 1 ml of 10x DMEM/F-12 medium to 8 ml Cellmatrix Type I-A and stir gently.
2. Add 1 ml of preparation buffer and stir gently.
3. Incubate the mixture at 4 °C for 30 min to remove any air bubbles, if necessary.
4. Store at 4 °C until use.
Procedure
1. Sterilize the MEA before the coating with collagen and perform all following steps under sterile
conditions.
2. Incubate the MEA at 4 °C for at least 1h.
3. Fill the MEA with collagen solution until the bottom of the culture chamber is completely covered.
Immediately remove the collagen solution with a glass pipette. The solution can be reused.
4. Incubate the MEA in a CO 2 incubator for 30 min.
5. Rinse the MEA with sterile distilled water.
6. Fill the MEA with culture medium and keep it sterile in a CO 2 incubator until use (for up to one
week).
7. Check for contaminations before use.
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