Session Name: Concurrent Session: Tissue Engineering & Technology
Date: Sunday, June 2, 2019
Session Time: 2:30pm-4:00pm
Presentation Time: 3:06pm-3:18pm
Location: Room 309
*Purpose: Hepatic stellate cells (HSC) comprise about 2-8% of liver cells and are vital to hepatocellular function. Scaffold-free 3D-bioprinting (SF3DBP) allows creation of realistic tissue models without the use of synthetic biomaterials. We hypothesize that co-culturing primary hepatocytes with HSCs and liver derived endothelial cells (LEC) will improve hepatocyte function and accuracy of the SF3DBP liver model.
*Methods: We used primary pig hepatocytes and immortalized pig HSCs and LECs to generate spheroids with hepatocytes alone, HSCs alone, or a combination of hepatocytes, HSCs, and LECs. Optimized combination spheroids were printed using a Regenova 3D-bioprinter. Un-printed spheroids were incubated over two weeks for functionality assays (albumin secretion, mRNA transcription, urea clearance) and immunostaining performed for cell composition confirmation.
*Results: Hepatocyte:HSC co-cultures (H/H, 2.5:1 ratio), Hepatocyte:HSC:LEC co-cultures (H/H/L 2.5:0.5:0.5 ratio) and HSC-only cells formed round spheroids within 48 hours, whereas the hepatocyte-only condition did not. Functional assays showed superiority in maintenance of albumin secretion and increased urea clearance over 14 days in H/H and H/H/L spheroids compared to hepatocyte culture alone. Real-Time PCR and immunostaining further confirmed that HSCs sustained the expression of hepatocyte marker gene expression (Alb and CK-18). After 4 days of spheroid incubation, optimized H/H combination spheroids were bioprinted. This SF3DBP construct fused 3 days post-bioprinting and could be removed from temporary microneedle support by 6 days post-bioprinting.
*Conclusions: SF3DBP of hepatocyte:HSC spheroids in a 2.5:1 ratio demonstrates the potential utility bioprinting has for pharmacological, immunological, and hepatotoxicity testing. Further spheroid optimization using different cell ratios including HSCs, hepatocytes, liver sinusoidal endothelial cells, and cholangiocytes will allow printing of more physiologically accurate liver models.
To cite this abstract in AMA style:Chen AM, Zhang W, Koehler CJ, Saglam K, Isidan A, Walsh JR, Naqvi RA, Gramelspacher E, Smith LJ, Li P, Ekser B. Hepatic Stellate Cells and Liver Derived Endothelial Cells Improve Scaffold-Free 3D-Bioprintied Liver Model [abstract]. Am J Transplant. 2019; 19 (suppl 3). https://atcmeetingabstracts.com/abstract/hepatic-stellate-cells-and-liver-derived-endothelial-cells-improve-scaffold-free-3d-bioprintied-liver-model/. Accessed July 24, 2021.
« Back to 2019 American Transplant Congress