| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Chemicals, peptides, and recombinant proteins | ||
| PBS (no calcium, no magnesium) | Thermo Fisher Scientific | Cat# 10010-023 |
| DMEM [+] 4.5 g/L glucose, sodium pyruvate [−] L-glutamine | VWR | Cat# 45000-316 |
| Gentamicin | GIBCO | Cat# 15750045 |
| Insulin, human recombinant, zinc solution | Thermo Fisher Scientific | Cat# 12585014 |
| Apo-transferrin | Sigma | Cat# T2252-500MG |
| HBSS (no calcium, no magnesium) | Thermo Fisher Scientific | Cat# 14175095 |
| Advanced DMEM/F12 | Thermo Fisher Scientific | Cat# 12634010 |
| GlutaMAX supplement | Thermo Fisher Scientific | Cat# 35050061 |
| Fetal bovine serum | Omega Scientific | Cat# FB-01 |
| HEPES | Thermo Fisher Scientific | Cat# 12587001 |
| Trypsin-EDTA (0.25%), phenol red | GIBCO | Cat# 25200056 |
| Cultrex basement membrane matrix, type2 (BME) | Trevigen | Cat# 3532-001-02 |
| B27 supplement minus vitamin A | Thermo Fisher Scientific | Cat# 12587001 |
| Murine EGF | Thermo Fisher Scientific | Cat# PMG8045 |
| Cell recovery solution | Corning | Cat# 354253 |
| Trypan blue solution | Gibco | Cat# 15250061 |
| Experimental models: cell lines | ||
| Mouse tumor organoids (MTOs)a | (Tauriello et al., 2018) | N/A |
| Mouse colonic fibroblasts (CFs) | (Kasashima et al., 2020) | N/A |
| Experimental models: organisms/strains | ||
| Mouse: WT (C57BL/6)b | Moscat lab | N/A |
| Other | ||
| Insulin syringe, 0.3 mL with 31-gauge needle | Becton Dickinson | Cat# 328438 |
| 15 mL polypropylene centrifuge tubes | Genesee Scientific | Cat# 28-103 |
| Bright-field microscope | EVOS M5000 | Cat# AMF5000 |
| Caliper | Mitutoyo | Cat# 500-171-30 |
| Scale (capable of measuring mouse weight) | Ohaus | Cat# SP202 |
| Sterile gauze | N/A | N/A |
| Electric small animal heated pad | K&H PET PRODUCTS | N/A |
| Isoflurane | VETONE | Vet-Rx-MW 502017 |
| Curved fine forceps | FST | Cat# 11159-10 |
| Straight brunt forceps | FST | Cat# 11002-12 |
| Straight scissors | FST | Cat# 14040-10 |
| Hartman hemostats | FST | Cat# 13002-10 |
| Autoclip applier 9-mm autoclips | Kent Scientific | Cat# INS750345; Cat# INS75045 |
| PERMA-HAND silk 5-0 | VWR | Cat# 95057-064 |
| Germinator 500 | Braintree Scientific | Cat# GER 5287-120V |
| Surgical microscope | Leica | N/A |
| Wound clip remover | FST | Cat#: 12033-00 |
a
This protocol can be used with other colorectal cell lines, such as MC-38 cells.
b
This protocol can be used in immunocompromised mice such as NSG mice if tumor producing rate is low.