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Table 1 Key features of methods described to prepare primary epithelial cell cultures from rectal biopsies of adult cattle

From: Lactoferrin translocates to the nucleus of bovine rectal epithelial cells in the presence of Escherichia coli O157:H7

  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%
  1. aThe method described by Mahajan et al. [26] was identical as the one used by Dziva et al. [25].