Multichannel Systems MEA Manual - Manual (Page 42)

MEA Manual
42
Procedure
1. Cover the MEA surface with 300 μl fibronectin solution and incubate the MEA at 37 °C
for at least 1 h.
2. Aspirate the solution and rinse the MEA 2 x with PBS (phosphate buffered saline).
3. Plate the cells onto the MEA immediately after coating.
Literature
Ulrich Egert, Thomas Meyer (2004); Heart on a Chip — Extracellular multielectrode recordings from
cardiac myocytes in vitro, "Methods in Cardiovascular Research", S. Dhein and M. Delmar (eds.)
5.4.7 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 (Gibco/Invitrogen, 21331-020)
 N Hydrochloric acid, pH 3.0
 Acid-soluble type I collagen solution (3 mg/ml, pH 3.0) Cellmatrix Type I-A (Nitta Gelatin Inc.)
Preparation buffer
 200 mM HEPES in 0.08 N NaOH
Collagen solution
 Add 1 ml of 10 x DMEM/F-12 medium to 8 ml Cellmatrix Type I-A and stir gently.
 Add 1 ml of preparation buffer and stir gently.
 Incubate the mixture at 4 °C for 30 min to remove any air bubbles, if necessary.
 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. Rinse the MEA with sterile distilled water. Fill the
MEA with culture medium and keep it sterile in a CO
2
incubator until use (for up to one week).
Check for contaminations before use.