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

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