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12日間培養したHeaptoCellsは、元の肝細胞の形態的特徴を維持し、CYP3A4 、1A2 や2B6 のCYP誘導活性といった典型的な初代肝細胞の特徴を維持していました。

画像: Corning HepatoCells を BioCoat コラーゲンI コート24ウェルプレート上で HepatoCells 培養培地にて培養した際の細胞画像。同様の形態は BioCoat コラーゲンIコート48ウェル、96ウェルでも観察された。

Phenobarbital-responsive CAR nuclear translocation, a phenomenon found in primary hepatocytes, was observed in Corning HepatoCells.  This indicates that the nuclear receptor pathways that are responsible for regulating the induction of CYP2B genes are intact in Corning HepatoCells. 

Image: EYFP-hCAR expression and localization in Corning HepatoCells and primary human hepatocytes (Data courtesy of Prof. Hongbing Wang, University of Maryland

成功事例のための具体的な情報は、このHepatoCellsユーザーガイドに記載された実験に必要なもの、凍結HepatoCellsの解凍方法、CYP450 誘導アッセイを行うためのプロトコールを参照ください。

HepatoCells were evaluated for their applicability as an in vitro screening tool for predicting clinical CYP3A4 inducers. Three lots of cells were treated with a group of known clinical strong, moderate/weak inducers, and non-inducers at more than 8 concentrations each. Results showed that all three lots of HepatoCells behaved similarly to primary human hepatocytes in terms of prediction, with only minor lot-to-lot variations for the 3 lots of cells.

Corning 96-well Ultra-Low Attachment spheroid microplates were used to culture
HepatoCells into single spheroids at indicated plating cell density (100mL/well). As shown in these photos taken on day 7, HepatoCells can form spheroids with cell number dependent size.

The performance of HepatoCells is compared to the parental hepatocyte cells and other hepatic models in order to determine if these cryopreserved cells model the behavior of parental primary human hepatocytes following in vitro exposure to known hepatotoxic compounds. Results suggest that HepatoCells closely model the functional behavior of parental cells and therefore may be used as a primary hepatocyte surrogate to study pharmacogenomic-related cytotoxicity.