MiCAN Technologies
cMylc
highly optimised myeloid cells from human iPSC suitable for SARS-CoV-2 vaccine and drug development
Details
Human blood cell based host cells to infect SAR-CoV-2 coronavirus
Ideal cell tool to evaluate ADE (antibody-dependent enhancement) for safe vaccine development
High reproducible cells using established stem cell technologies
SARS-CoV-2 Infection Data
SARS-CoV-2 virus successfully infected cMylc cells expressing ACE2 and TMPRSS2. cMylc-DC cells, further differentiated using the cMylc-kit are more susceptible to SARS-CoV-2 virus.
Viral RNA detection from supernatant of the infected cell culture. Significant increase of progeny viruses were observed on cMylc-DC with further differentiation.
Viral RNA detection from supernatant of the infected cell culture. Significant increase of progeny viruses were observed on cMylc-DC with further differentiation.
SARS-CoV-2 Infection Data
SARS-CoV-2 virus successfully infected cMylc cells expressing ACE2 and TMPRSS2. cMylc-DC cells, further differentiated using the cMylc-kit are more susceptible to SARS-CoV-2 virus.
Viral RNA detection from supernatant of the infected cell culture. Significant increase of progeny viruses were observed on cMylc-DC with further differentiation.
Viral RNA detection from supernatant of the infected cell culture. Significant increase of progeny viruses were observed on cMylc-DC with further differentiation.
Experimental flow using "cMylc"
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.
[Full protocol is available upon purchasing and request]
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.
"cMylc" is provided with supplement media as a kit
cMylc cryopreserved cell vial
Storage: Frozen, under -150 degC
Shipping: Frozen, dry-ice
Supplement media A&B for cMylc cells
Storage: Frozen, under -80 degC (recommended)
Shipping: Frozen, dry-ice