$MCET Granted Patent Drug: Multicell Technologies Immortalized Hepatocytes Doc Type Granted Patent in US, Europe and Germany!
Immortalized Human Hepatocyte Cell Lines [0084]In another preferred embodiment of the present invention, the cell lines proliferate easily in media. Preferably, the cell lines proliferate easily in a serum-free media. More preferably, the cell lines proliferate easily in MFE media (MultiCell Technologies Inc., Providence, R.I., USA; XenoTech, LLC, Lenexa, Kans., USA). https://www.lens.org/lens/patent/167-627-709-236-287/fulltext
INTRODUCTION Primary cultures of human hepatocytes are widely used to evaluate the cytochrome P450 (CYP) enzyme-inducing potential and/or toxicity of drug candidates. However, the availability of human hepatocytes for this purpose is limited and erratic, and there are large inter-individual differences in the magnitude of induction (due to differences in both "control" and "induced" CYP activities). Immortalized human hepatocytes, Fa2N-4, developed by Multicell Technologies, (Warwick, RI) have the potential to overcome these limitations. The cells, immortalized through transformation of human hepatocytes with SV40 T antigen, display in vitro cell morphology that closely resembles primary human hepatocytes (Figure 1). The Fa2N-4 cells retain many of the characteristics of primary hepatocytes, including the inducibility of multiple CYP mRNAs and enzyme activities (Mills et al., 2002; Morris et al., 2003; Czerwinski et al., 2003). In this study, we further characterized the ability of these cells to respond to enzymeinducing xenobiotics. Additionally, we examined the toxicity profile of multiple enzyme inducers in primary hepatocytes and Fa2N-4 cells.
MATERIALS AND METHODS The Fa2N-4 cells were propagated in MFE media (Multicell Technologies) on plasticware coated with Vitrogen (Cohesion Technologies, Palo Alto, CA) and maintained at 37°C, 5% CO2, 95% humidity. Immortalized hepatocytes were grown to confluency in 24 or 96-well plates and treated with enzyme inducers for up to 6 consecutive days with daily changes of medium. All enzyme inducers were dissolved in DMSO (final concentration of solvent 0.1%, v/v). Primary human hepatocytes were isolated and cultured as described (LeCluyse et al., 2000, Madan et al., 2003). Enzymatic activities were determined by incubating the cells with CYPspecific substrates. Metabolite formation was measured by LC-MS as summarized in Table 1.
Application US10/574,163 events 2003-10-10 Priority to US51050903P 2004-10-07 Application filed by Multicell Technologies Inc 2007-01-04 Publication of US20070004039A1 2009-07-28 Publication of US7566567B2 2009-07-28 Application granted Status Active 2025-01-13 Adjusted expiration https://patents.google.com/patent/US7566567B2/un
EP 1704227 B1 Doc Type: Granted Patent ID: lens.org/097-040-197-694-101 Jan 27, 2010
EP1704227B1 European Patent Office
Application EP04794437A events 2003-10-10 Priority to US51050903P 2004-10-07 Application filed by Multicell Technologies Inc 2004-10-07 Priority to PCT/US2004/033091 2006-09-27 Publication of EP1704227A1 2007-01-17 Publication of EP1704227A4 2010-01-27 Application granted 2010-01-27 Publication of EP1704227B1 Status Active 2024-10-07 Anticipated expiration https://patents.google.com/patent/EP1704227B1/en
DE 602004025380 D1 Doc Type: Granted Patent ID: lens.org/082-791-344-403-259 Mar 18, 2010
Application DE602004025380T events 2003-10-10 Priority to US51050903P 2004-10-07 Application filed by Multicell Technologies Inc 2004-10-07 Priority to PCT/US2004/033091 2010-03-18 Publication of DE602004025380D1 Status Active 2024-10-08 Anticipated expiration https://patents.google.com/patent/DE602004025380/D1
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