Presentation Information
[2ACCE-03]Engineered Hepatic Fibers for Off-the-Shelf Bioartificial Liver and Toxicity Screening Application
○Silas Habimana1, Lucas Trindade2, Nana Shirakigawa1, Gigante Ian Paulo Calicdan1, Hiroyuki Kitano1, Yoshinori Kawabe1, Masamichi Kamihira1,3 (1. Kyushu Univ. (Japan), 2. CellFiber Co., Ltd. (Japan), 3. Correspondence (Japan))
Keywords:
heat-inducible hepatic cells,hydrogel fiber encapsulation
Hepatic models that combine scalability with functional fidelity are critical for bioartificial liver development and hepatotoxicity testing. Primary human hepatocytes are limited by donor variability and rapid functional decline, while iPSC-derived hepatocytes often exhibit immature phenotypes and require costly differentiation protocols, highlighting the need for alternative cell sources.
We established heat-inducible hepatic cells (hi-Hep), a HepG2-derived cell line engineered with eight liver-enriched transcription factors (LETFs) and BAP1 under the control of a heat-shock protein promoter. Transient heat induction (43 oC, 30 min) rapidly activated hepatic functions. Transcriptomic profiling by DNA microarray was performed under both monolayer and spheroid culture conditions. hi-Hep cells were encapsulated in core-shell hydrogel fibers using double-coaxial microfluidic extrusion and validated in a lab-scale circulation tissue culture circuit. To enable true off-the-shelf deployment, we further developed a cryobanking strategy that preserves cell viability, structural fiber integrity, and inducible hepatic functionality after-thawing.
Heat induction activated albumin secretion, ammonia detoxification, cytochrome P450 activity, and bile acid metabolism. Transcriptomic profiling confirmed activation of liver-specific gene programs and revealed a maturation state superior to that of both parental HepG2 cells and HepG2/8F_HS cells, with key hepatic markers (ALB, ARG1, CPS1, CYP3A4, CYP7A1, CYP27A1) approaching or matching levels observed in primary human hepatocytes under serum-free culture conditions. In the fiber configuration, ammonia clearance reached 62%, albumin secretion reached 47 pg cell-1 day-1, and cell viability remained at 98% during a 24-hour perfusion culture. Cryopreserved fiber constructs rapidly recovered metabolic activity following heat-induction, demonstrating suitability for storage, transport, and on-demand clinical and pharmaceutical applications.
The integration of genetic programming, fiber-based 3D culture, and cryobanking establishes hi-Hep fibers as a scalable, reproducible, and ready-to-use platform for bioartificial liver devices and standardized drug toxicity screening.
We established heat-inducible hepatic cells (hi-Hep), a HepG2-derived cell line engineered with eight liver-enriched transcription factors (LETFs) and BAP1 under the control of a heat-shock protein promoter. Transient heat induction (43 oC, 30 min) rapidly activated hepatic functions. Transcriptomic profiling by DNA microarray was performed under both monolayer and spheroid culture conditions. hi-Hep cells were encapsulated in core-shell hydrogel fibers using double-coaxial microfluidic extrusion and validated in a lab-scale circulation tissue culture circuit. To enable true off-the-shelf deployment, we further developed a cryobanking strategy that preserves cell viability, structural fiber integrity, and inducible hepatic functionality after-thawing.
Heat induction activated albumin secretion, ammonia detoxification, cytochrome P450 activity, and bile acid metabolism. Transcriptomic profiling confirmed activation of liver-specific gene programs and revealed a maturation state superior to that of both parental HepG2 cells and HepG2/8F_HS cells, with key hepatic markers (ALB, ARG1, CPS1, CYP3A4, CYP7A1, CYP27A1) approaching or matching levels observed in primary human hepatocytes under serum-free culture conditions. In the fiber configuration, ammonia clearance reached 62%, albumin secretion reached 47 pg cell-1 day-1, and cell viability remained at 98% during a 24-hour perfusion culture. Cryopreserved fiber constructs rapidly recovered metabolic activity following heat-induction, demonstrating suitability for storage, transport, and on-demand clinical and pharmaceutical applications.
The integration of genetic programming, fiber-based 3D culture, and cryobanking establishes hi-Hep fibers as a scalable, reproducible, and ready-to-use platform for bioartificial liver devices and standardized drug toxicity screening.
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