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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

cMylc infection data 1.tif

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.

cMylc infection data 3.jpg

Viral RNA detection from supernatant of the infected cell culture. Significant increase of progeny viruses were observed on cMylc-DC with further differentiation.

cMylc infection data 2.tif

SARS-CoV-2 Infection Data

cMylc infection data 1.tif

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.

cMylc infection data 2.tif

Viral RNA detection from supernatant of the infected cell culture. Significant increase of progeny viruses were observed on cMylc-DC with further differentiation.

cMylc infection data 3.jpg

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"

cMylc flow.jpg

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

mMylc frozen vial image.png

cMylc cryopreserved cell vial

Storage: Frozen, under -150 degC

Shipping: Frozen, dry-ice

Supplement A.png
Supplement B.png

Supplement media A&B for cMylc cells

Storage: Frozen, under -80 degC (recommended)

Shipping: Frozen, dry-ice

For collaboration request