Open Access
Table 1
Guidelines for construction of collagen I (1.5 mg/mL) based dermal layer.
| Constituents | Calculated vol. of each constituent (µL) |
Calculated vol. of each constituent (µL) |
|---|---|---|
| Total vol. of acellular layer = 200 µL per insert | Total vol. of cellular layer = 400 µL per insert | |
| 10× M199 (Sigma) | 20 µL (10% of final vol. of 200 µL results in 1× M199) | 40 µL (10% of final vol. of 400 µL results in 1× M199) |
| 200 mM Glutamine (Biosciences) | 0.68 µL | 1.36 µL |
| Formula: M1·V1 = M2·V2 (200 mM × V1 = 0.68 × 200 µL) (This 0.68 µL vol. results in 0.68 mM Glutamine in final 200 µL vol. of acellular layer) | Formula: M1·V1 = M2·V2 (200 mM × V1 = 0.68 × 400 µL) (This 1.36 µL vol. results in 0.68 mM Glutamine in final 400 µL vol. of cellular layer) | |
| ddH2O | 113.32 µL (vol. calculated in the end) | 176.64 µL (vol. calculated in the end) |
| 7.5% of NaHCO3 (Sigma) | 6 µL (This volume is 10% of the volume of collagen I, that results in a final mixture pH of 7.2–7.4 – optimized) | 12 µL (This is an optimized volume as 10% of the volume of collagen I, that results in a final mixture pH of 7.2–7.4) |
| 5 mg/mL Collagen I (Ibidi) | 60 µL (This volume makes final conc. of 1.5 mg/mL in final volume) | 120 µL (This volume makes final conc. of 1.5 mg/mL in final volume) |
| NHDF (PromoCell) | N/A | 50 µL (this volume should contain 5 × 104 NHDF per insert) |
| Total vol. | 200 µL | 400 µL |