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Long-Term Culture and Coculture of Primary Rat and Human Hepatocytes
Open accessAbstract
The liver is the largest internal organ in mammals, serving a wide spectrum of vital functions. Loss of liver function due to drug toxicity or viral infection is a major cause of death in the United States. The development of Bioartificial Liver (BAL) devices and the demand for pharmaceutical and cosmetic toxicity screening require the development of long-term hepatocyte culture techniques. However, primary hepatocytes rapidly lose their cuboidal morphology and liver-specific functions over a few days in culture. Accumulation of stress fibers, loss of metabolic function, and cell death are known phenomena. In recent years, several techniques were developed that can support high levels of liver-specific gene expression, metabolic and synthetic function for several weeks in culture. These include the collagen double-gel configuration, hepatocyte spheroids, coculture with endothelial cells, and micropatterned cocultures with 3T3-J2 fibroblasts. This chapter covers the current status of hepatocyte culture techniques, including: hepatocyte isolation, media formulation, oxygen supply, heterotypic cell-cell interactions, and basic functional assays.
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“However, the limited proliferative capacity of PHs, often associated with reduced viability and progressive loss of hepatocyte-specific functions in vitro, has constrained their broader contribution to health and science [39].”
From The Hepatocyte Expansion Paradox: A Review of In Vitro Challenges and Advances (2026) · cites this paper as [39]
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Amino acid levels determine metabolism and CYP450 function of hepatocytes and hepatoma cell lines
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Human Nonalcoholic Steatohepatitis on a Chip
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“Elafibranor is a dual agonist for peroxisome proliferator-activated receptors (PPAR-α/δ), which are highly involved in regulation of fatty acid metabolism in HCs.”
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“In addition, the phenotype is unstable and primary cells can only be cultured for a short time span [3,4].”
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Moya, A., Ortega‐Ribera, M., Villalba, X. G., Sowade, E., Zea, M., Illa, X., Ramón, E., Villa, R., Gracia‐Sancho, J., & Gabriel, G. (2018) · Lab on a Chip · doi:10.1039/c8lc00456k · Cited by 150
Osteoprotegerin Promotes Liver Steatosis by Targeting the ERK–PPAR-γ–CD36 Pathway
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“Cell Culture and Treatment Mouse primary hepatocytes (MPHs) were isolated from WT, OPG, and CD36 mice aged 8 weeks, as previously reported (31).”
MethodAvior, Y., Levy, G., Zimerman, M., Kitsberg, D., Schwartz, R., Sadeh, R., Moussaieff, A., Cohen, M., Itskovitz‐Eldor, J., & Nahmias, Y. (2015) · Hepatology · doi:10.1002/hep.27803 · Cited by 88
Self-citationCalcium Signaling in Liver Injury and Regeneration
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“Furthermore, PHH in 2D culture form actin stress fibers, resulting in fibroblast-like cell morphology (43).”
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“Further, in standard 2D cultures, primary hepatocytes show a rapid loss of the original phenotype, resulting in the downregulation of metabolic biomarkers (Shulman & Nahmias, 2013).”
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Human Induced Pluripotent Stem Cells in Hepatology
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Huang, D. (., Gibeley, S. B., Xu, C., Xiao, Y., Celik, O., Ginsberg, H. N., & Leong, K. W. (2020) · Advanced Functional Materials · doi:10.1002/adfm.201909553 · Cited by 65
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“[1] This limits or prevents their use for clinical, engineering and research purposes.”
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“Primary rat hepatocytes were isolated by two-step collagenase digestion following isolation of the liver from male Sprague Dawley® rats (Charles River Laboratories, Seattle, WA) (Shulman and Nahmias, 2013).”
MethodBioengineering Organs for Blood Detoxification
Legallais, C., Kim, D., Mihaila, S. M., Mihajlovic, M., Figliuzzi, M., Bonandrini, B., Salerno, S., Yengej, F. A. Y., Rookmaaker, M. B., Romero, N. S., et al. (2018) · Advanced Healthcare Materials · doi:10.1002/adhm.201800430 · Cited by 67
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In vitro and in silico liver models: Current trends, challenges and opportunities
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“One of the most promising approaches to overcome the issues with viability of hepatocyte monoculture is the utilization of microfluidic perfusion devices (Knöspel et al., 2016; Shulman and Nahmias, 2013; Tehranirokh et al., 2013; Wagner et al., 2013).”
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Human hepatocyte systems for in vitro toxicology analysis
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“It is thus not surprising that pHH performance turned better upon co-culturing with endothelial cells [24, 25] and through formation of 3D organizations in spheroids [26] or by employing micro-patterned structures [27].”
BackgroundPeptide-based scaffolds for the culture and maintenance of primary human hepatocytes
MacPherson, D., Bram, Y., Park, J., & Schwartz, R. E. (2021) · Scientific Reports · doi:10.1038/s41598-021-86016-5 · Cited by 36
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Grossini, E., Bellofatto, K., Farruggio, S., Sigaudo, L., Marotta, P., Raina, G., de Giuli, V., Mary, D., Pollesello, P., Minisini, R., et al. (2015) · PLoS ONE · doi:10.1371/journal.pone.0124742 · Cited by 30
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MethodIsolation, culture, and functional analysis of hepatocytes from mice with fatty liver disease
Jung, Y., Zhao, M., & Svensson, K. J. (2020) · STAR Protocols · doi:10.1016/j.xpro.2020.100222 · Cited by 42
Narkhede, A. A., Shevde, L. A., & Rao, S. S. (2017) · International Journal of Cancer · doi:10.1002/ijc.30748 · Cited by 36
“A major drawback of direct co-culture is its inability to maintain the metabolic ability of hepatocytes for a long time.(72) This makes it difficult to study the chemoresistance of BCCs in a physiologically relevant hepatic niche.”
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Sherlock, L. G., Sjostrom, K., Lei, S., Delaney, C. A., Tipple, T. E., Krebs, N. F., Nozik‐Grayck, E., & Wright, C. J. (2020) · Frontiers in Immunology · doi:10.3389/fimmu.2020.595282 · Cited by 27
“Isolation of Primary Hepatocytes Primary hepatocytes were isolated as previously described (36, 37).”
MethodWei, G., Wang, J., Lv, Q., Liu, M., Xu, H., Zhang, H., Jin, L., Yu, J., & Wang, X. (2018) · Journal of Biomedical Materials Research Part A · doi:10.1002/jbm.a.36421 · Cited by 32
DOCK5 regulates energy balance and hepatic insulin sensitivity by targeting mTORC1 signaling
Lai, Y., Zhao, A., Tan, M., Yang, M., Lin, Y., Li, S., Song, J., Zheng, H., Zhu, Z., Liu, D., et al. (2019) · EMBO Reports · doi:10.15252/embr.201949473 · Cited by 32
“MPHs were isolated from DOCK5 (cid:1) / (cid:1) , Raptor flox/flox , or WT mice aged 8 weeks as previously reported [47].”
Hoshi, M., Osawa, Y., Nakamoto, K., Morita, N., Yamamoto, Y., Ando, T., Tashita, C., Nabeshima, T., & Saito, K. (2020) · Toxicology · doi:10.1016/j.tox.2020.152458 · Cited by 28
Nguyen, R., Bae, S. D. W., Zhou, G., Read, S. A., Ahlenstiel, G., George, J., & Qiao, L. (2020) · Biochimica et Biophysica Acta (BBA) - Reviews on Cancer · doi:10.1016/j.bbcan.2020.188350 · Cited by 27
Nanoparticle Drug Delivery Can Reduce the Hepatotoxicity of Therapeutic Cargo
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“However, in vitro cultivation of PHHs often faces obstacles including failure of long-term in vitro culture, rapid loss of liver-specific function and morphology, tendency of fibrosis, and weak proliferation ability.(7,8) Besides, the extensive use of cancerous liver cell lines, e.”
BackgroundMethodCreating rat hepatocyte organoid as an in vitro model for drug testing
He, Y.-T., Zhu, X.-L., Li, S.-F., Zhang, B.-Q., Li, Y., Wu, Q., Zhang, Y.-L., Zhou, Y.-Y., Li, L., Qi, Y.-N., et al. (2020) · World Journal of Stem Cells · doi:10.4252/wjsc.v12.i10.1184 · Cited by 22
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“However, the practical application of primary hepatocytes has been frustrated by their tendency to rapidly dedifferentiate and lose most hepatic functions after growth in a tissue culture environment [1–4].”
BackgroundChida, T., Ishida, Y., Morioka, S., Sugahara, G., Han, C., Lam, B., Yamasaki, C., Sugahara, R., Li, M., Tanaka, Y., et al. (2023) · JCI Insight · doi:10.1172/jci.insight.162404 · Cited by 18
“To develop a comprehensive understanding of the host response to HDV infection, the use of a physiological experimental platform, terminally differentiated HH, is essential; however, this has been also regarded as a research tool with limited versatility because of a rapid loss of genuine characteristics during in vit…”
BackgroundHuman Liver-Derived Extracellular Matrix for the Culture of Distinct Human Primary Liver Cells
Alevra Sarika, N., Payen, V. L., Fléron, M., Ravau, J., Brusa, D., Najimi, M., De Pauw, E., Eppe, G., Mazzucchelli, G., Sokal, E. M., et al. (2020) · Cells · doi:10.3390/cells9061357 · Cited by 18
“It is indeed well-known that, upon in vitro culture, HEPs rapidly lose their polarity and metabolic properties [7,8], HSCs undergo an activation process during which they rapidly lose their retinoid-containing lipid droplets and acquire a fibrogenic, myofibroblast-like phenotype [9,10], and LSECs lose their fenestrati…”
BackgroundSani, F., Zomorrod, M. S., Azarpira, N., & Soleimani, M. (2023) · Stem Cells International · doi:10.1155/2023/8836452 · Cited by 19
“It has been determined that hepatocytes, HSCs, and endothelial cells make up 60 % – 80 % , 20 % , and 15 % – 20 % , respectively, of the total cells in the natural human liver tissue [14 – 17].”
RetractedIsolation and cultivation of adult primary bovine hepatocytes from abattoir derived liver
Ehrhardt, S., & Schmicke, M. (2016) · EXCLI journal · doi:10.17179/excli2016-794 · Cited by 14
“Primary hepatocytes obtained from adult mice and rats are widely used for toxicological and metabolic studies and well standardized (Hewitt et al., 2007; Shulman and Nahmias, 2013).”
MethodLiver organoids: From fabrication to application in liver diseases
Liu, Q., Zeng, A., Liu, Z., Wu, C., & Song, L. (2022) · Frontiers in Physiology · doi:10.3389/fphys.2022.956244 · Cited by 19
“Two-dimensional cultured cell models, such as PHHs and induced pluripotent stem cells (IPSCs) also lose the polarity that hepatocytes exhibit in vivo because they are forced to adopt a flat morphology in vitro (Shulman and Nahmias, 2013).”
BackgroundHepG2 and human healthy hepatocyte in vitro culture and co-culture in PCL electrospun platforms
Fasolino, I., Guarino, V., Marrese, M., Cirillo, V., Vallifuoco, M., Tamma, M., Vassallo, V., Bracco, A., Calise, F., & Ambrosio, L. (2017) · Biomedical Materials · doi:10.1088/1748-605x/aa8c51 · Cited by 20
Impact of Percoll purification on isolation of primary human hepatocytes
Horner, R., Gassner, J. G. M. V., Kluge, M., Tang, P., Lippert, S., Hillebrandt, K. H., Moosburner, S., Reutzel‐Selke, A., Pratschke, J., Sauer, I. M., et al. (2019) · Scientific Reports · doi:10.1038/s41598-019-43042-8 · Cited by 19
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“The use of the collagen sandwich culture system provides conditions to ensure well-differentiated phenotype of hepatocytes and to reestablish cell polarity [42] as was seen in the present study with cultures of hepatocytes being well-differentiated and possibly even forming bile canaliculi, as observed previously [43].”
ResultKhalil, A., Al-Massarani, G., Aljapawe, A., Ekhtiar, A. M., & Bakir, M. A. (2020) · Frontiers in Pharmacology · doi:10.3389/fphar.2020.528400 · Cited by 13
Cell Culture Systems for Studying Hepatitis B and Hepatitis D Virus Infections
Lee, S.-H., Purdy, M. A., & Choi, Y. H. (2023) · Life · doi:10.3390/life13071527 · Cited by 14
“The most common 3D culture method is placing PHHs on a single-layer collagen matrix; however, basic liver functions decrease in this cell culture system after a week [152].”
Simple Paper-based Liver Cell Model for Drug Screening
Kaarj, K., Ngo, J., Loera, C., Akarapipad, P., Cho, S., & Yoon, J.-Y. (2020) · BioChip Journal · doi:10.1007/s13206-020-4211-6 · Cited by 11
Thasler, W. E., Thasler, R., Schelcher, C., & Jauch, K. (2013) · Langenbeck s Archives of Surgery · doi:10.1007/s00423-013-1060-y · Cited by 11
“In some cases, experimental set-up requires primary human cells to stay in culture over a long period of time and since hepatocytes have a relatively short life span in culture, co-culturing with a variety of non-parenchymal cells such as Kupffer, stellate and epithelial cells is necessary in order to increase their l…”
BackgroundWu, Y. C., Wu, G. X., Chen, K., Shiu, L., Kumar, S., Liu, G., & Kuo, S. M. (2022) · Scientific Reports · doi:10.1038/s41598-022-05174-2 · Cited by 13
“The description of pathological features: (1) reticular fibers circumscribing the nodules; (2) obvious formation of pseudolobules; (3) numerous fibrocytes; (4) hepatocyte necrosis; (5) swollen hepatocytes; (6) large/moderate decreases in fibrous tissues/fibrocytes; (7) proliferation of the mini-bile duct; (8) fatty de…”
BackgroundIntegrative microphysiological tissue systems of cancer metastasis to the liver
Clark, A. M., Allbritton, N. L., & Wells, A. H. (2020) · Seminars in Cancer Biology · doi:10.1016/j.semcancer.2020.06.010 · Cited by 11
3D Co-Culture with Vascular Cells Supports Long-Term Hepatocyte Phenotype and Function In Vitro
Unal, A. Z., Jeffs, S. E., & West, J. L. (2018) · Regenerative Engineering and Translational Medicine · doi:10.1007/s40883-018-0046-2 · Cited by 12
Manuguerra, S., Carli, F., Scoditti, E., Santulli, A., Gastaldelli, A., & Messina, C. M. (2024) · Metabolites · doi:10.3390/metabo14100559 · Cited by 8
“Moreover, hepatoma cells overcome the limitations of using primary hepatocytes, mostly in screening studies, including the limited availability of liver samples, inter-individual variability, limited survival, loss of metabolic activity after long culture, and costs [76,77].”
BackgroundLASS2 regulates hepatocyte steatosis by interacting with NDUFS2/OXPHOS related proteins.
Yang, Y., Yang, X.-L., Lin, Y., Yang, G.-Y., & Li, L. (2020) · Biochemical and Biophysical Research Communications - BBRC · doi:10.1016/j.bbrc.2020.02.166 · Cited by 8
Huang, T.-Y., Wang, G.-S., Ko, C.-S., Chen, X.-W., & Su, W.-T. (2020) · Materials Science and Engineering C: Materials for Biological Applications · doi:10.1016/j.msec.2019.110563 · Cited by 8
Radko, L., Śniegocki, T., Sell, B., & Posyniak, A. (2019) · Animals · doi:10.3390/ani10010030 · Cited by 7
“Primary cultures of hepatocytes and liver-derived lines are the models that are used for the evaluation of drugs of which toxicity is mediated through biotransformation [22,23].”
BackgroundTafti, Z. A., Mahmoodi, M., Hajizadeh, M. R., Ezzatizadeh, V., Baharvand, H., Vosough, M., & Piryaei, A. (2018) · Europe PMC (PubMed Central) · doi:10.22074/cellj.2018.5288 · Cited by 11
Naito, Y., Yoshinouchi, Y., Sorayama, Y., Kohara, H., Kitano, S., Irie, S., & Matsusaki, M. (2021) · Acta Biomaterialia · doi:10.2139/ssrn.3906842 · Cited by 7
A three‐dimensional microfluidized liver system to assess hepatic drug metabolism and hepatotoxicity
Corrado, B., Gregorio, V. D., Imparato, G., Attanasio, C., Urciuolo, F., & Netti, P. A. (2018) · Biotechnology and Bioengineering · doi:10.1002/bit.26902 · Cited by 34
Ullah, I., Kim, Y., Lim, M.-U., Oh, K. B., Hwang, S., Shin, Y., Kim, Y.-I., Im, G., Hur, T., & Ock, S. A. (2017) · Animal Cells and Systems · doi:10.1080/19768354.2017.1381151 · Cited by 5
“D EV EL O PM EN TA L BI O LO G Y CONTACT Sun-A Ock ouksuna@hotmail.com; ocksa@korea.kr Animal Biotechnology Division, National Institute of Animal Science, RDA, 1500 Kongjwipatjwi-ro Iseo-myeon, Wanju-gun, Jeollabuk-do 565-851, Republic of Korea (Shulman and Nahmias 2013).”
BackgroundInfluentialFroghi, S., de Andrade, M. O., Mohammad-Hadi, L., Gélat, P., Rashidi, H., Quaglia, A., Fuller, B. J., Saffari, N., & Davidson, B. R. (2023) · Bioengineering · doi:10.3390/bioengineering10020276 · Cited by 6
Liu, H., You, S., Rong, Y., Wu, Y., Zhu, B., Zhihong, W., Liu, W., Panyong, M., & Xin, S. (2013) · Human Cell · doi:10.1007/s13577-013-0068-5 · Cited by 7
“The elimination of glucose was mainly related with energy needs in culturing time [24, 25].”
Status and Future of 3D Cell Culture in Toxicity Testing
Otieno, M. A., Gan, J., & Proctor, W. R. (2018) · Methods in pharmacology and toxicology · doi:10.1007/978-1-4939-7677-5_12 · Cited by 6
Kim, H. M., Kim, Y., Kim, Y., Kim, Y. J., & Ko, K. S. (2023) · Preventive Nutrition and Food Science · doi:10.3746/pnf.2023.28.3.360 · Cited by 6
“Furthermore, primary hepatocytes rapidly lose their morphological characteristics (Shulman and Nahmias, 2012; Rose et al., 2021).”
BackgroundSerrano, L. J., García-Arranz, M., Pablo-Moreno, J. A. D., Segovia, J., Olivera-Salazar, R., García-Olmo, D., & Liras, A. (2022) · International Journal of Molecular Sciences · doi:10.3390/ijms23105802 · Cited by 5
Александровна, Ш. И., Prozorova, G. F., Сергеевна, Т. И., Korzhova, S. A., Дремина, Н. Н., Emel’yanov, A. I., Say, O., Kuznetsova, N. P., Pozdnyakov, A. S., & Шурыгин, М. Г. (2021) · Nanomaterials · doi:10.3390/nano12010016 · Cited by 4
“The cells were used for the experiment within 24 h after isolation since no changes in the hepatocyte enzymatic systems functioning were observed during this period [53,54].”
BackgroundMethodManipulating the mechanics of extracellular matrix to study effects on the nucleus and its structure
Xia, Y., Cho, S., Vashisth, M., Ivanovska, I. L., Dingal, P. C. D. P., & Discher, D. E. (2018) · Methods · doi:10.1016/j.ymeth.2018.12.009 · Cited by 5
“Ignoring the diversity of mechanical cues presented to cells in normal and diseased tissues surrenders control of additional factors that affect cell and nuclear phenotype [4,69].”
Ravichandran, S., & del Sol, A. (2017) · FEBS Letters · doi:10.1002/1873-3468.12559 · Cited by 7
Morita, A., Yamada, M., Utoh, R., Momiyama, K., Iwadate, H., & Seki, M. (2021) · Journal of Bioscience and Bioengineering · doi:10.1016/j.jbiosc.2021.11.009 · Cited by 5
Tafti, Z. A., Mahmoodi, M., Hajizadeh, M., Ezzatizadeh, V., Baharvand, H., Massoud Vosough, M. P., & Piryaei, A. (2021) · Cell journal · doi:10.22074/cellj.2021.7936 · Cited by 4
Functional Liver Cell–Based Platforms in Biomedical Research
Hashemian, Z., Taleahmad, S., Shokouhian, B., Najimi, M., & Vosough, M. (2025) · Pharmacology Research & Perspectives · doi:10.1002/prp2.70128 · Cited by 2
“This progress enabled researchers to monitor drug metabolism and distribution within the body, providing a proper platform for drug pharmacokinetics studies [62–65].”
Weber, M., Linti, C., Lörch, C., Weber, M., Andt, M., Schlensak, C., Wendel, H. P., Doser, M., & Avci‐Adali, M. (2023) · Scientific Reports · doi:10.1038/s41598-023-49117-x · Cited by 7
“Primary hepatocytes rapidly lose their liver-specific functions, including CYP inducibility and the production of plasma proteins like albumin, during in vitro cultivation 39 .”
Functional characterization of a bovine luteal cell culture model: Effects of passage number
Sharma, A., Vanselow, J., Kühn, C., & Becker, D. (2025) · PLoS ONE · doi:10.1371/journal.pone.0334047 · Cited by 2
“Highly proliferative cells of organs such as kidney [7], liver [8] and pancreas [9] have been long-term cultured but lose their cell-type specific functions in vitro.”
Yee, C., Chan, Y. L., Utama, R. H., Besnier, M., Engel, M., & Belfiore, L. (2024) · STAR Protocols · doi:10.1016/j.xpro.2024.103234 · Cited by 2
Fan, J. Y., Dama, G., Liu, Y. L., Guo, W. Y., & Lin, J. T. (2023) · Molecular Biology · doi:10.1134/s0026893323040039 · Cited by 2
“However, primary hepatocytes exhibit a restricted ability to proliferate, with a decrease in cell viability and loss of hepatocyte-specific functions often observed in vitro [22].”
BackgroundNeo, S., MAKIISHI, E., Fujimoto, A., & Hisasue, M. (2020) · Journal of Veterinary Medical Science · doi:10.1292/jvms.20-0320 · Cited by 3
“Long-term hepatocyte culture techniques are also required to establish artificial hepatocytes for pharmaceutical toxicity screening [25].”
BackgroundLa plasticidad del hepatocito y su relevancia en la fisiología y la patología hepática
Ríos-López, D. G., Aranda-López, Y., Sosa‐Garrocho, M., & Macías‐Silva, M. (2020) · TIP Revista Especializada en Ciencias Químico-Biológicas · doi:10.22201/fesz.23958723e.2020.0.225 · Cited by 4
Chinnici, C., Miceli, V., Pampalone, M., Nigro, A. L., Amico, G., & Conaldi, P. (2017) · Biochemical and Biophysical Research Communications - BBRC · doi:10.1016/j.bbrc.2017.06.065 · Cited by 2
One step antibody-mediated isolation and patterning of multiple cell types in microfluidic devices.
Bavli, D., Ezra, E., Kitsberg, D., Vosk-Artzi, M., Murthy, S., & Nahmias, Y. (2016) · Biomicrofluidics · doi:10.1063/1.4944741 · Cited by 2
Self-citationPorcine Hepatocytes: Isolation and Liver Tissue Engineering for Xenotransplantation
Li, Y., Wu, Q., Wang, Y., Bu, H., & Bao, J. (2020) · Methods in molecular biology · doi:10.1007/978-1-0716-0255-3_18 · Cited by 4
Reference Gene Validation for Quantitative PCR Analysis in 2D and 3D AML12 Hepatocyte Models
Ivanova, Z., Petrova, V., Todorova, B., Penev, T. G., & Grigorova, N. (2026) · Biomedicines · doi:10.3390/biomedicines14010150 · Cited by 2
Predicting Systemic and Liver Bosentan Exposure Using Physiologically‐Based Pharmacokinetic Modeling
Huang, M., Macente, J., Heylen, S., Chen, N., Vos, K. D., Deferm, N., & Annaert, P. (2025) · CPT Pharmacometrics & Systems Pharmacology · doi:10.1002/psp4.70085 · Cited by 1
“However, it is known that primary hepatocytes dedifferentiate and exhibit reduced expression of enzymes and transporters over the culture time [35, 36], and show low induction sensitivity for drug metabolic enzymes like CYP2Cs [37].”
Sibiya, T., Xaba, L., Mthethwa, L., Chuturgoon, A. A., & Msomi, N. (2025) · Viruses · doi:10.3390/v17081057 · Cited by 2
“One method involves placing PHHs on a single-layer collagen matrix; however, this results in a decline in basic liver functions [124].”
Proposal for a non-adhesive single-cell culture technology for primary hepatocytes
Uehara, M. K., Bual, R. P., Shafiq, M., Yoshida, K., & Ijima, H. (2024) · Cytotechnology · doi:10.1007/s10616-024-00696-1 · Cited by 1
“This morphology is much closer to the native cuboidal morphology of PHs within the liver (Shulman and Nahmias 2012b). mRNA expression levels for some typical liver-function related genes (Alb, Cps1, Cyp1a1, Hnf4a, etc.) at 24 h showed no reduction in the PHs treated with PEG-GRGDS compared to those…”
InfluentialIsolation and Cryopreservation of Primary Macaque Hepatocytes
Guo, X.-Q., Seo, J.-E., Davis, K., Malhi, P., & Fili, C. (2024) · Current Protocols · doi:10.1002/cpz1.70015 · Cited by 1
“This is considered the best time-frame for performing the experiment because primary hepatocytes rapidly lose their hepatic features over a few days in culture (Shulman & Nahmias, 2013).”
Fatty Liver Disease and Utility of Stem Cells in Developing the Disease Model
Choudhary, S., LaCasse, M., Beitz, D. C., & Testroet, E. D. (2021) · Springer eBooks · doi:10.1007/978-981-16-3464-2_6
Kim, Y.-H., Özer, S., & Uygun, B. E. (2013) · Elsevier eBooks · doi:10.1016/b978-0-12-398523-1.00024-0 · Cited by 1
The Hepatocyte Expansion Paradox: A Review of In Vitro Challenges and Advances
Kolahdouzmohammadi, M., Tjandra, N., Wu, K., Nikoumaram, R., & Oldani, G. (2026) · Cells · doi:10.3390/cells15161465
“However, the limited proliferative capacity of PHs, often associated with reduced viability and progressive loss of hepatocyte-specific functions in vitro, has constrained their broader contribution to health and science [39].”
Tran, H. V. A., Han, S.-H., Tran, T. X. T., Kim, K. W., Kim, M. H., Lee, I.-J., & Kwak, J.-Y. (2026) · International Journal of Molecular Sciences · doi:10.3390/ijms27052366
Appendix A: Drug Metabolism in Drug Discovery
Coleman, M. D. (2020) · Wiley eBooks · doi:10.1002/9781119658016.app1
A34 EFFECT OF AUTOPHAGY INDUCTION VIA TRPML1 ACTIVATION ON HEPATIC STEATOSIS
Tersigni, C., Capurro, M., MacParland, S. A., Selzner-Malekkiani, N., & Jones, N. L. (2019) · Journal of the Canadian Association of Gastroenterology · doi:10.1093/jcag/gwz006.033
Cholesterol metabolism and statin effects on an FH class II LDL-receptor mutation.
Omer, L. (2018) · doi:10.18297/etd/3080
“Culturing and expanding primary hepatocytes has proven difficult since their survival depends on the liver microenvironment [271, 272].”
Background
Metadata
- Authors
- Shulman, M., & Nahmias, Y.
- Affiliations
- Hebrew University of Jerusalem (IL)
- Corresponding author
- Nahmias, Yaakov
- ORCID
- Nahmias, Yaakov
- Venue
- Methods in Molecular Biology
- Publisher
- Humana Press
- Published
- 2012
- Volume
- 945
- Pages
- 287-302
- ISSN
- 1064-3745 (ISSN-L)
- 1940-6029
- PMC
- PMC3781339
- OpenAlex
- W1850526299
- Cited by
- 117
- References
- 52
- Citations by year
- 2026: 1 · 2025: 5 · 2024: 6 · 2023: 8 · 2022: 9 · 2021: 8 · 2020: 19 · 2019: 9 · 2018: 18 · 2017: 13 · 2016: 6 · 2015: 6 · 2014: 2 · 2013: 7
- FWCI
- 15.92
- Type
- book chapter
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- Locations
- Methods in molecular biology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute of Diabetes and Digestive and Kidney Diseases (K01DK080241)
- Topics
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Keywords
- primary hepatocytes, long-term hepatocyte culture, hepatocyte co-culture, bioartificial liver, hepatocyte spheroids, hepatocyte isolation, culture medium formulation, oxygen supply, hepatocyte death, drug toxicity testing
- MeSH terms
- Albumins, Animals, Cell Separation, Enzyme-Linked Immunosorbent Assay, Humans, Oxygen, Perfusion, Time Factors, Urea, Coculture Techniques, Cell Culture Techniques, Hepatocytes, Rats
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