Coronavirus optimised Myeloid lineage cells (human iPSC derived) with designated cMylc culture media (supplement A and B) as a kit.
MiCAN - cMylc (coronavirus optimised myeloid lineage cell) kit
[Full protocol is available upon purchasing and request]
1. Preparation of culture medium
50 uL of supplement is added into 10 mL 10% FBS/aMEM. Remaining supplement could be kept at 4 degC for two weeks (refreezing and thawing not recommended).
2. Preparation of cMylc cells
Cryopreserved cMylc in a frozon vial are thawed rapidly in a 37 degC water bath, and immediately suspended in 10% FBS/aMEM at room temperature.
3. Cell culture of cMylc
After centrifuging (300G, 5 min at room temperature), cell pellets are resuspended in 10 mL of 10% FBS/aMEM for two days in a humidified atomosphere under 5% CO2 (95% air) at 37 degC.
After two days, cells are centributed at above condition. Cell pellets are resuspended in 10 mL of 10% FBS/aMEM indluding "supplement B" and incubated for 3 days at 37 degC in a low cell binding T-25 flask. cMylc-DC, which is ideal for virus infection, is ready for experiment.
4. Viris infection
cMylc-DC is prepared in a 96 well cell culture plate to be mixed with SARS-CoV-2 viruses. 4 hours after mixing, the inoculum is completely removed from each well, then 10% FBS/aMEM is added. Progeny viruses are detectable in the supernatant after 3 days with quantitative RT-PCR.