Hepatic microphysiological system: Tumor co-culture and drug response

The ex vivo hepatic MPS (Legacy LiverChip^® by CNBio Innovations Ltd.) was assembled, seeded and maintained as previously described (19 (link), 26 ). The functioning and bioengineering behind the MPS (e.g. physiological mimicry, fluid flow and oxygenation etc.) are also been explained in detail elsewhere (28 (link), 29 (link)). Briefly, hepatocytes and non-parenchymal cells were seeded onto scaffolds coated with 1% rat-tail collagen type I (BD Biosciences) at a 1:1 ratio (6x10⁵ cells/scaffold) in William’s E Medium (WE; Life Technologies) supplemented with the Hepatocyte Thawing and Plating Supplement Pack (Life Technologies). Cells were cultured overnight and then the medium was changed to WE supplemented with the Hepatocyte Maintenance Supplement Pack (Life Technologies). After allowing the hepatic tissue to form, MDA-MB-231 cells expressing RFP (500 cells/scaffold) were introduced on day 3. Applicable cultures were treated with 1 µM doxorubicin (APP Pharmaceuticals LLC) on day 7 to 10 and then stimulated on day 13 to 15. The stimulus of 0.5, 1.0 or 5.0 ng/mL IP-10 (PeproTech) was administered in the presence or absence of 50 nM AMG-487 (Tocris). On day 15, scaffolds were removed and fixed in 2% paraformaldehyde at 4°C for 1 hour.

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Clark A.M., Heusey H.L., Griffith L.G., Lauffenburger D.A, & Wells A. (2021). IP-10 (CXCL10) Can Trigger Emergence of Dormant Breast Cancer Cells in a Metastatic Liver Microenvironment. Frontiers in Oncology, 11, 676135.

Publication 2021

rat-tail collagen type I Hepatocyte Thawing and Plating Supplement Pack Hepatocyte Maintenance Supplement Pack IP-10 AMG-487

Amg 487 Cells Collagen type i Doxorubicin Hepatocyte Innovations Mda mb 231 cells Oxygenation Paraformaldehyde Pharmaceuticals Physiological Supplement Tail Tissue

Corresponding Organization : McGowan Institute for Regenerative Medicine

Other organizations : Massachusetts Institute of Technology

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Variable analysis

independent variables

Concentration of IP-10 (0.5, 1.0 or 5.0 ng/mL)
Presence or absence of 50 nM AMG-487

dependent variables

Response of MDA-MB-231 cells expressing RFP to the treatments

control variables

Hepatocytes and non-parenchymal cells seeded onto scaffolds coated with 1% rat-tail collagen type I at a 1:1 ratio (6x10^5 cells/scaffold)
Cell culture medium (William's E Medium supplemented with Hepatocyte Thawing and Plating Supplement Pack, and Hepatocyte Maintenance Supplement Pack)
Treatment with 1 µM doxorubicin from day 7 to 10
Introduction of MDA-MB-231 cells expressing RFP (500 cells/scaffold) on day 3

positive controls

Applicable cultures treated with 1 µM doxorubicin on day 7 to 10

negative controls

Cultures treated with IP-10 in the absence of 50 nM AMG-487

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