In vitro culture of isolated primary hepatocytes and stem cell-derived hepatocyte-like cells for liver regeneration

Protein & Cell, Jun 2015

Various liver diseases result in terminal hepatic failure, and liver transplantation, cell transplantation and artificial liver support systems are emerging as effective therapies for severe hepatic disease. However, all of these treatments are limited by organ or cell resources, so developing a sufficient number of functional hepatocytes for liver regeneration is a priority. Liver regeneration is a complex process regulated by growth factors (GFs), cytokines, transcription factors (TFs), hormones, oxidative stress products, metabolic networks, and microRNA. It is well-known that the function of isolated primary hepatocytes is hard to maintain; when cultured in vitro, these cells readily undergo dedifferentiation, causing them to lose hepatocyte function. For this reason, most studies focus on inducing stem cells, such as embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), hepatic progenitor cells (HPCs), and mesenchymal stem cells (MSCs), to differentiate into hepatocyte-like cells (HLCs) in vitro. In this review, we mainly focus on the nature of the liver regeneration process and discuss how to maintain and enhance in vitro hepatic function of isolated primary hepatocytes or stem cell-derived HLCs for liver regeneration. In this way, hepatocytes or HLCs may be applied for clinical use for the treatment of terminal liver diseases and may prolong the survival time of patients in the near future.

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In vitro culture of isolated primary hepatocytes and stem cell-derived hepatocyte-like cells for liver regeneration

© The Author(s) 2015. This article is published with open access at Springerlink.com and journal.hep.com.cn In vitro culture of isolated primary hepatocytes and stem cell-derived hepatocyte-like cells for liver regeneration Chenxia Hu 0 Lanjuan Li 0 0 Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, School of Medicine, First Affiliated Hospital, Zhejiang University , Hangzhou 310006 , China liver regeneration; primary hepatocyte; stem cell; hepatocyte-like cell; in vitro culture - Various liver diseases result in terminal hepatic failure, and liver transplantation, cell transplantation and artificial liver support systems are emerging as effective therapies for severe hepatic disease. However, all of these treatments are limited by organ or cell resources, so developing a sufficient number of functional hepatocytes for liver regeneration is a priority. Liver regeneration is a complex process regulated by growth factors (GFs), cytokines, transcription factors (TFs), hormones, oxidative stress products, metabolic networks, and microRNA. It is well-known that the function of isolated primary hepatocytes is hard to maintain; when cultured in vitro, these cells readily undergo dedifferentiation, causing them to lose hepatocyte function. For this reason, most studies focus on inducing stem cells, such as embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), hepatic progenitor cells (HPCs), and mesenchymal stem cells (MSCs), to differentiate into hepatocyte-like cells (HLCs) in vitro. In this review, we mainly focus on the nature of the liver regeneration process and discuss how to maintain and enhance in vitro hepatic function of isolated primary hepatocytes or stem cell-derived HLCs for liver regeneration. In this way, hepatocytes or HLCs may be applied for clinical use for the treatment of terminal liver diseases and may prolong the survival time of patients in the near future. Viral hepatitis, fatty liver disease, drug-induced liver injury, liver cirrhosis, hepatic carcinoma, and other liver diseases can cause acute or chronic liver failure. Approximately 10% of patients with liver disease succumb to their condition while waiting for liver sources each year (Kim et al., 2006). Liver transplantation was once the only therapeutic option for patients with end stage liver diseases, and its clinical use was limited due to limited donor availability, surgical injuries, a high incidence of surgical complications and the high cost of the treatment (Duan et al., 2013). Later, cell transplantation and artificial liver support emerged as effective methods for compensation of lost liver function and increased the survival rate of patients; however, these two methods are also limited by the availability of effective cell sources and equipment. The inability of hepatocytes to proliferate in vitro and the severely inadequate supply of hepatocytes due to donor shortage are still the main problems for primary human hepatocyte-based treatments. Stem cells have been proposed as an ideal cell source because they have potent selfrenewal, low immunogenicity, and the capacity to differentiate into various cell types. Furthermore, they can generate unlimited hepatocytes with incomplete function (Sancho-Bru et al., 2009) that are generally defined as hepatocyte-like cells (HLCs). HLCs can be derived from multiple stem cell types, such as embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), hepatic progenitor cells (HPCs), and mesenchymal stem cells (MSCs). Therefore, it is crucial to develop robust methods for differentiating stem cells into mature hepatocytes in vitro for clinical use. Here, we present an overview of isolated primary hepatocytes and stem cell-derived HLCs used for liver regeneration and describe how the in vitro environment in which they are cultured is continuously being optimized to mimic in vivo conditions and maintain hepatic function. The main disadvantages, histologic origin, 3D, and co-culture environment for in vitro culture of isolated hepatocytes or stem cellderived hepatocytes were demonstrated in Table 1. Optimization of in vitro culturing of functional hepatocytes will solve the issues of limited cell numbers and limited function, and sufficient numbers of functional hepatocytes will be used to promote liver regeneration directly or indirectly. NATURE OF LIVER REGENERATION The liver serves as a major storage site of glycogen and vitamin A and is one of only a few organs in adults that are capable of regeneration. Normal mature hepatocytes and cholangiocytes stay in the G0 phase of the cell cycle, exhibit a quiescent phenotype and show minimal turnover, but in response to partial hepatectomy (PH), they can undergo cell proliferation to compensate for the lost cells, a process called liver regeneration. However, severe damage caused by liver diseases can signif (...truncated)


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Chenxia Hu, Lanjuan Li. In vitro culture of isolated primary hepatocytes and stem cell-derived hepatocyte-like cells for liver regeneration, Protein & Cell, 2015, pp. 562-574, Volume 6, Issue 8, DOI: 10.1007/s13238-015-0180-2