Table 1 Key features of methods described to prepare primary epithelial cell cultures from rectal biopsies of adult cattle
Dziva et al. [25]a | Sheng et al. [27] | Present study | |
|---|---|---|---|
Enzymatic disintegration | Collagenase 75 U/mL Dispase 20 mg/mL | Collagenase 75 U/mL Accutase 20 mg/mL | Collagenase 100 U/mL |
Mechanical disintegration | No | No | Yes |
Differential centrifugation | Yes | Yes | Yes |
Insulin | 0.25 U/mL | 0.25 U/mL | 0.25 U/mL |
Epidermal growth factor | 10 ng/mL | 10 ng/mL | 10 ng/mL |
FCS | 2.5% | 2.5% | 10% during first 24 h Afterwards 1% |
Number of crypts planted | 400–600 | 400 | 200–250 |
Coating with collagen | Yes | Yes | Yes |
Fibroblast growth inhibition | d-Valine | Cis-OH-proline | Repeated gentle trypsinization Reduction of FCS to 1% |