Table 1 Pig intestinal organoids

Duodenum

Lipopolysaccharides challenge

30 mM EDTA, 1.5 mM DTT, 4 °C, 20 min + 30 mM EDTA, 10 µM Y27632, warm, 10 min

DMEM, N2, B27 without vitamin A, 10 mM HEPES, 2 mM L-glutamine, 1 μg/mL R-spondin1, 100 ng/mL Noggin, 50 ng/mL EGF, 2.5 ng/mL Wnt-3a

[60]

Gene edited pigs

1 mg/mL collagenase, 2 mg/mL BSA, 37 °C, 30 min

IntestiCult™ (STEMCELL Technologies, Vancouver, Canada)

[37]

Jejunnum

Cell lineage identification

30 mM EDTA, 1 mM DTT, 10 µM Y27632, 4 °C, 20 min and 37 °C 10 min

DMEM/F12, N-2, B-27 minus vitamin A, Glutamax, 1 mM HEPES, 1 mM NAC, 100 ng/mL rm Noggin, 500 ng/mL rh R-spondin, 50 ng/mL rm EGF, 100 ng/mL rh Wnt3a, 10 µM Y-27632, 10 µM SB202190, 500 nM LY2157299

[39]

Lentivirus transduction

10 mM EDTA, 1 mM DTT, 4 °C, 30 min

DMEM/F-12, 2 mM glutaMAX, 10 mM HEPES, 1 mM NAC, N2, B27, 50 ng/mL rm EGF, 100 ng/mL rm Noggin, 500 ng/mL rh R-spondin 1, Wnt3a CM, 1 mM valproic acid, 10 mM nicotinamide, 2.5 μM CHIR99021, 10 μM Y27632, 10 μM SB202190, 500 nM LY2157299

[25]

2D monolayers, transcriptomics

2 mM EDTA, 4 °C, 30 min

DMEM/F12, 10 mM HEPES, B-27, 1.25 mM NAC, 50 ng/mL rh EGF, 15 nM gastrin, 10 mM nicotinamide, 10 μM p38 MAPK inhibitor, 600 nM A83-01, Noggin CM (15% v/v), R-spondin CM (15% v/v), and Wnt3A CM (30% v/v)

[5, 19]

Mycotoxin challenge

30 mM EDTA, 1.5 mM DTT, 10 µM Y27632, 4 °C, 30 min

DMEM, 10% FBS, N2, B27, 1X GlutaMAX, 1 mM NAC, 50 ng/mL EGF, 100 ng/mL Noggin, 500 ng/mL R-spondin1, 10 mM nicotinamide, 10 µM SB202190, 0.5 µM LY2157299, 45% Wnt3a CM, 10 µM Y27632 (2–3 first days)

[18]

Heat Stress

30 mM EDTA, 1.5 mM DTT, 4 °C, 30 min

DMEM/F12, Wnt3a CM, rm EGF, rm Noggin, rh R-spondin1, nicotinamide, Y27632, LY2157299, SB202190, CHIR99021, FBS, N2, B27, glutamine, NAC

[26]

Swine enteric virus infection

5 mM EDTA, 4 °C, 1 h

IntestiCult™ (STEMCELL Technologies, Vancouver, Canada)

[34]

S. Typhimurium and T. gondii infection

0.8 mM EDTA, 4 °C, 30 min

IntestiCult™ (STEMCELL Technologies, Vancouver, Canada)

[20]

Ileum

Model development

Collagenase, 37 °C, 1.5 h

50% (DMEM + 10% FBS), 50% L-WRN CM, 10 µM SB431542, 10 µM Y27632

[14]

Vitamin A

2 mM EDTA, 4 °C, 30 min

Wnt3a, Noggin and R-spondin 1 CM, B27, N2, glutamine, NAC, rm EGF, nicotinamide, SB202190

[45]

Mucus production

30 mM EDTA, 1 mM DTT, 37 °C, 10–15 min

IntestiCult™ (STEMCELL Technologies, Vancouver, Canada)

[43]

Lawsonia intracellularis infection

30 mM EDTA, 1 mM DTT, 37 °C, 5–15 min

IntestiCult™ (STEMCELL Technologies, Vancouver, Canada)

[36]

Duodenum, Jejunum, Ileum, Colon

Porcine epidemic diarrhea virus infection

Cell dissociation reagent (STEMCELL Technologies,Vancouver, Canada)

IntestiCult™ (STEMCELL Technologies, Vancouver, Canada)

[21]