3D Spheroid Culture for HBV Infection

3D spheroid cultures were performed as previously described^{18 (link)}. Cells were seeded into 96-well ultra-low attachment plates (Corning, Amsterdam, The Netherlands) at a density of 1500 cells/well and were maintained in WEM supplemented with Thawing/Plating Supplement Pack (Invitrogen) for 5 days prior to HBV infection until the spheroid formation is readily detectable by microscopy. Medium was subsequently replaced with WEM supplemented with Cell Maintenance Supplement Pack (Invitrogen) and 50% of the media volume was replaced every 48 h.

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Ortega-Prieto A.M., Skelton J.K., Wai S.N., Large E., Lussignol M., Vizcay-Barrena G., Hughes D., Fleck R.A., Thursz M., Catanese M.T, & Dorner M. (2018). 3D microfluidic liver cultures as a physiological preclinical tool for hepatitis B virus infection. Nature Communications, 9, 682.

Publication 2018

96-well ultra-low attachment plates Thawing/Plating Supplement Pack Cell Maintenance Supplement Pack

Cell Infection Microscopy Supplement

Corresponding Organization :

Other organizations : Imperial College London, CN Bio Innovations (United Kingdom), King's College London

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Variable analysis

independent variables

Seeding density of cells (1500 cells/well)

dependent variables

Spheroid formation

control variables

96-well ultra-low attachment plates (Corning, Amsterdam, The Netherlands)
WEM supplemented with Thawing/Plating Supplement Pack (Invitrogen) for 5 days prior to HBV infection
WEM supplemented with Cell Maintenance Supplement Pack (Invitrogen) after spheroid formation
Medium replacement every 48 h (50% of the media volume)

controls

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

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