Maintenance of Human Hepatocyte Function in Vitro by Liver-derived Extracellular Matrix Gels

Overview

Journal Tissue Eng Part A

Specialties Anatomy & Histology
Biomedical Engineering
Biotechnology

Date 2009 Oct 23

PMID 19845461

Citations 113

Authors

Tiffany L Sellaro

Aarati Ranade

Denver M Faulk

George P McCabe

Kenneth Dorko

Stephen F Badylak

Stephen C Strom

Affiliations

Soon will be listed here.

Abstract

Tissue engineering and regenerative medicine (TE&RM) approaches to treating liver disease have the potential to provide temporary support with biohybrid-liver-assist devices or long-term therapy by replacing the diseased liver with functional constructs. A rate-limiting step for TE&RM strategies has been the loss of hepatocyte-specific functions after hepatocytes are isolated from their highly specialized in vivo microenvironment and placed in in vitro culture systems. The identification of a biologic substrate that can maintain a functional hepatocyte differentiation profile during in vitro culture would advance potential TE&RM therapeutic strategies. The present study compared two different biologic substrates for their ability to support human hepatocyte function in vitro: porcine-liver-derived extracellular matrix (PLECM) or Matrigel. Because Matrigel has been shown to be the most useful matrix for static, traditional hepatocyte culture, we directly compared PLECM with Matrigel in each experiment. Albumin secretion, hepatic transport activity, and ammonia metabolism were used to determine hepatocyte function. Hepatocytes cultured between two layers of PLECM or Matrigel showed equally high levels of albumin expression and secretion, ammonia metabolism, and hepatic transporter expression and function. We conclude that like Matrigel, PLECM represents a favorable substrate for in vitro culture of human hepatocytes.

Citing Articles

3D bioprinting technology innovation in female reproductive system.

Chen S, Wang T, Chen J, Sui M, Wang L, Zhao X Mater Today Bio. 2025; 31:101551.

PMID: 40026632 PMC: 11870202. DOI: 10.1016/j.mtbio.2025.101551.

Application of Additive Manufacturing in Assisted Reproductive Techniques: What Is the Evidence? A Clinical and Technical Systematic Review of the Literature.

Kontogeorgi A, Boutas I, Tsangkalova G, Messaropoulos P, Koufopoulos N, Schwab R Medicina (Kaunas). 2024; 60(11).

PMID: 39597074 PMC: 11596948. DOI: 10.3390/medicina60111889.

Advances in xenogeneic donor decellularized organs: A review on studies with sheep and porcine-derived heart valves.

Inal M, Avci H, Hassan S, Darcan C, Shin S, Akpek A Bioeng Transl Med. 2024; 9(6):e10695.

PMID: 39545084 PMC: 11558188. DOI: 10.1002/btm2.10695.

Advanced biomaterials for regenerative medicine and their possible therapeutic significance in treating COVID-19: a critical overview.

Sarangi A, Salem M, Younus M, El-Haroun H, Mahal A, Tripathy L Int J Surg. 2024; 110(12):7508-7527.

PMID: 39411890 PMC: 11634172. DOI: 10.1097/JS9.0000000000002110.

An Ultrathin Coating of Microcapsules Enhances the Function of Encapsulated Hepatocyte Spheroids.

Choi D, Gwon K, de Hoyos-Vega J, Lee S, Nguyen K, Gonzalez-Suarez A ACS Appl Mater Interfaces. 2024; 16(38):51411-51420.

PMID: 39269915 PMC: 11702863. DOI: 10.1021/acsami.4c08329.

References

Zeisberg M, Kramer K, Sindhi N, Sarkar P, Upton M, Kalluri R . De-differentiation of primary human hepatocytes depends on the composition of specialized liver basement membrane. Mol Cell Biochem. 2006; 283(1-2):181-9. DOI: 10.1007/s11010-006-2677-8. View

Dunn J, Tompkins R, Yarmush M . Long-term in vitro function of adult hepatocytes in a collagen sandwich configuration. Biotechnol Prog. 1991; 7(3):237-45. DOI: 10.1021/bp00009a007. View

Strom S, Pisarov L, Dorko K, Thompson M, Schuetz J, Schuetz E . Use of human hepatocytes to study P450 gene induction. Methods Enzymol. 1996; 272:388-401. DOI: 10.1016/s0076-6879(96)72044-x. View

Sidhu J, Farin F, Omiecinski C . Influence of extracellular matrix overlay on phenobarbital-mediated induction of CYP2B1, 2B2, and 3A1 genes in primary adult rat hepatocyte culture. Arch Biochem Biophys. 1993; 301(1):103-13. DOI: 10.1006/abbi.1993.1121. View

Reid L, Gaitmaitan Z, Arias I, Ponce P, Rojkind M . Long-term cultures of normal rat hepatocytes on liver biomatrix. Ann N Y Acad Sci. 1980; 349:70-6. DOI: 10.1111/j.1749-6632.1980.tb29516.x. View

Moghe P, Berthiaume F, Ezzell R, Toner M, Tompkins R, Yarmush M . Culture matrix configuration and composition in the maintenance of hepatocyte polarity and function. Biomaterials. 1996; 17(3):373-85. DOI: 10.1016/0142-9612(96)85576-1. View

Lecluyse E, Madan A, Hamilton G, Carroll K, DeHaan R, Parkinson A . Expression and regulation of cytochrome P450 enzymes in primary cultures of human hepatocytes. J Biochem Mol Toxicol. 2000; 14(4):177-88. DOI: 10.1002/(sici)1099-0461(2000)14:4<177::aid-jbt1>3.0.co;2-4. View

Gilbert T, Sellaro T, Badylak S . Decellularization of tissues and organs. Biomaterials. 2006; 27(19):3675-83. DOI: 10.1016/j.biomaterials.2006.02.014. View

Jadhav A, Mason H, Cepko C . Notch 1 inhibits photoreceptor production in the developing mammalian retina. Development. 2006; 133(5):913-23. DOI: 10.1242/dev.02245. View

10.

Jasmund I, Schwientek S, Acikgoz A, Langsch A, Machens H, Bader A . The influence of medium composition and matrix on long-term cultivation of primary porcine and human hepatocytes. Biomol Eng. 2006; 24(1):59-69. DOI: 10.1016/j.bioeng.2006.05.018. View

11.

Reid L, Fiorino A, Sigal S, Brill S, Holst P . Extracellular matrix gradients in the space of Disse: relevance to liver biology. Hepatology. 1992; 15(6):1198-203. DOI: 10.1002/hep.1840150635. View

12.

LeCluyse E, Audus K, Hochman J . Formation of extensive canalicular networks by rat hepatocytes cultured in collagen-sandwich configuration. Am J Physiol. 1994; 266(6 Pt 1):C1764-74. DOI: 10.1152/ajpcell.1994.266.6.C1764. View

13.

Michalopoulos G, Sattler G, Sattler C, Pitot H . Interaction of chemical carcinogens and drug-metabolizing enzymes in primary cultures of hepatic cells from the rat. Am J Pathol. 1976; 85(3):755-72. PMC: 2032658. View

14.

Lin P, Chan W, Badylak S, Bhatia S . Assessing porcine liver-derived biomatrix for hepatic tissue engineering. Tissue Eng. 2004; 10(7-8):1046-53. DOI: 10.1089/ten.2004.10.1046. View

15.

Schuetz E, Li D, Omiecinski C, Muller-Eberhard U, Kleinman H, Elswick B . Regulation of gene expression in adult rat hepatocytes cultured on a basement membrane matrix. J Cell Physiol. 1988; 134(3):309-23. DOI: 10.1002/jcp.1041340302. View

16.

Silva J, Morin P, Day S, Kennedy B, Payette P, Rushmore T . Refinement of an in vitro cell model for cytochrome P450 induction. Drug Metab Dispos. 1998; 26(5):490-6. View

17.

Kato S, Otsu K, Ohtake K, Kimura Y, Yashiro T, Suzuki T . Concurrent changes in sinusoidal expression of laminin and affinity of hepatocytes to laminin during rat liver regeneration. Exp Cell Res. 1992; 198(1):59-68. DOI: 10.1016/0014-4827(92)90149-3. View

18.

Cepko C . The roles of intrinsic and extrinsic cues and bHLH genes in the determination of retinal cell fates. Curr Opin Neurobiol. 1999; 9(1):37-46. DOI: 10.1016/s0959-4388(99)80005-1. View

19.

Futaki S, Hayashi Y, Yamashita M, Yagi K, Bono H, Hayashizaki Y . Molecular basis of constitutive production of basement membrane components. Gene expression profiles of Engelbreth-Holm-Swarm tumor and F9 embryonal carcinoma cells. J Biol Chem. 2003; 278(50):50691-701. DOI: 10.1074/jbc.M304985200. View

20.

Clark P, Connolly P, Curtis A, Dow J, Wilkinson C . Topographical control of cell behaviour: II. Multiple grooved substrata. Development. 1990; 108(4):635-44. DOI: 10.1242/dev.108.4.635. View