| MEM (Sigma-Aldrich M7278) |
|
394 mL |
| Heat-inactivated horse seruma |
20% |
100 mL |
| l-glutamine (200 mm; Gibco 25030-024)
|
1 mm |
2.5 mL |
| Insulin (1 mg/mL; Sigma-Aldrich I6634) |
0.01 mg/mL |
0.5 mL |
| NaCl (5 m; Sigma-Aldrich S5150)
|
|
1.45 mL |
| MgSO4 (1 m; Fluka 63126)
|
2 mm |
1 mL |
| CaCl2 (1 m; Fluka 21114)
|
1.44 mm |
0.72 mL |
| Ascorbic acid (25%; Fluka 11140) |
0.00125% |
2.4 µL |
| d-glucose (Fluka 49152)
|
13 mm |
1.16 g |
| aThe serum is often a critical factor in slice quality and it is often necessary to test several batches (lots); three products
that have been successfully used are Sigma-Aldrich H1138, Gibco 26050070, and Gibco 16050122. Gibco 16050122 must be heat-inactivated
for 30 min at 55°C.
|
| After mixing, filter-sterilize (0.2-µm pore size), and store at 4°C in 50-mL aliquots. (The solution should be orange-red
[i.e., pH ∼7.3] and osmolality should be ∼320 mOsm/kg.)
|
