Cardiac Organoid Manipulation for MMP14 Knockdown

Human cardiac organoids were purchased from Novoheart (Hong Kong) and cultured with StemPro34 SFM (Invitrogen) medium supplemented with ascorbic acid (AA, 50 mg/mL; Sigma), 2 mM GlutaMAX-1 (Invitrogen), BMP4 (10 ng/mL), IWR-1 (5 mM; Enzo Life Sciences) and human recombinant activin-A (10 ng/mL; Invitrogen). The culture medium was replaced with the StemPro34 SFM medium with AA every two days^{70 (link)}. Cardiac organoids were disassembled using TrypLE express upon reaching confluence, and infected with shMMP14 lentivirus for 24 h. Cardiac organoids were subsequently selected with Puromycin (0.5 ug/ml) for one week.

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Guo X., Cao J., Cai J.P., Wu J., Huang J., Asthana P., Wong S.K., Ye Z.W., Gurung S., Zhang Y., Wang S., Wang Z., Ge X., Kwan H.Y., Lyu A., Chan K.M., Wong N., Huang J., Zhou Z., Bian Z.X., Yuan S, & Wong H.L. (2022). Control of SARS-CoV-2 infection by MT1-MMP-mediated shedding of ACE2. Nature Communications, 13, 7907.

Publication 2022

StemPro34 SFM GlutaMAX-1 IWR-1 human recombinant activin-A

Ascorbic acid Bmp4 Cardiac Human Lentivirus Organoids Puromycin Recombinant activin a

Corresponding Organization :

Other organizations : Hong Kong Baptist University, University of Hong Kong, Chinese University of Hong Kong, Xiamen University, Guang Fu Hospital, The University of Texas Health Science Center at Houston, City University of Hong Kong, Prince of Wales Hospital

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Variable analysis

independent variables

ShMMP14 lentivirus infection

dependent variables

Cardiac organoid characteristics

control variables

Cardiac organoid culture conditions (StemPro34 SFM medium with ascorbic acid, GlutaMAX-1, BMP4, IWR-1, human recombinant activin-A)
Puromycin selection

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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