Table 1. Summary of preferred conditions found for co-culture.
| Factor | Preferred condition |
|---|---|
| Substrate material | Polystyrene (tissue-culture treated) |
| SMC growth medium | DMEM with 10% FBS, 10% PS, and growth factors (20% total serum) |
| SMC quiescent medium | Serum-free DMEM/F12 with ITS supplement |
| EC growth medium | Same as SMC growth medium except with 5% FBS, 5% PS (10% total serum) |
| Co-culture medium | 1 part growth medium: 5 parts quiescent medium (3.3% total serum) |
| SMC seeding density | 25 × 103−50 × 103 cells/cm2 a |
| EC seeding density | 40 × 103−80 × 103 cells/cm2 a |
| SMC phenotype | Quiescent (contractile) |
| Basal adhesion protein | FN (5 μg/cm2) |
| Medial adhesion protein | No difference observed between collagen IV, FN, and no protein in terms of EC coverage and PECAM expression |
a
The range for the seeding density reflects the fact that size of EC and SMC from different source varies. SMCs are seeded at a subconfluent density, whereas ECs are seeded at a confluent density. The optimal seeding density has to be determined in each batch of cells.