SteCM (Stellate Cell Medium)培养

星状细胞培养基是为正常人类肝脏星状细胞体外培养设计的适于其生长的培养基。经灭菌的液体培养基，包含必需和非必需氨基酸、维生素、有机和无机化合物、激素、生长因子、微量矿物质和低浓度胎牛血清(2%)。该培养基缓冲体系为重碳酸盐，在含5%CO2的细胞培养箱中平衡后pH值为7.4。该培养基的配方能够选择性的促进正常人类星形细胞体外培养中的增殖和生长，并为其达到理想营养平衡状态提供数量上和质量上的保证。

星状细胞培养基包含500 ml基础培养基，10ml胎牛血清(FBS,目录编号0010)，5ml星状细胞生长添加物(SteCGS,目录编号5352)和5 ml青霉素/链霉素溶液(P/S,目录编号0503)

1. Sasaki, R., Kanda, T., Nakamura, M., Nakamoto, S., Haga, Y., Wu, S., Shirasawa, H. & Yokosuka, O. (2016) Possible Involvement of Hepatitis B Virus Infection of Hepatocytes in the Attenuation of Apoptosis in Hepatic Stellate Cells PLoS One. 11

Background The induction of apoptosis in hepatic stellate cells (HSCs) is a promising therapeutic strategy against hepatitis B virus (HBV)-related hepatic fibrosis. The underlying mechanisms of apoptosis in HSCs, however, are unknown under consideration of HBV infection. In this study, the effects of HBV on apoptosis and endoplasmic reticulum (ER) stress signaling in HSCs were examined. Methods The effects of conditioned media (CM) from HepG2.2.15 on apoptosis induced by the proteasome inhibitor MG132 in LX-2 and HHSteC were studied in regard to c-Jun. In combination with c-Fos, c-Jun forms the AP-1 early response transcription factor, leading to AP-1 activation, signal transduction, endoplasmic reticulum (ER) stress and apoptosis. Results In LX-2 cells, MG132 treatment was associated with the phosphorylation of c-Jun, activation of AP-1 and apoptosis. However, in the presence of CM from HepG2.2.15, these phenomena were attenuated. In HHSteC cells, similar results were observed. HBV genomic DNA is not involved in the process of HSC apoptosis. It is possible that HBeAg has an inhibitory effect on MG132-induced apoptosis in LX-2. We also observed the upregulation of several ER stress-associated genes, such as cAMP responsive element binding protein 3-like 3, inhibin-beta A and solute carrier family 17-member 2, in the presence of CM from HepG2.2.15, or CM from PXB cells infected with HBV. Conclusions HBV inhibits the activation of c-Jun/AP-1 in HSCs, contributing to the attenuation of apoptosis and resulting in hepatic fibrosis. HBV also up-regulated several ER stress genes associated with cell growth and fibrosis. These mechanistic insights might shed new light on a treatment strategy for HBV-associated hepatic fibrosis. Less

2. Kramer, B., Finnemann, C., Sastre, B., Lutz, P., Gl?ssner, A., Wolter, F., Goeser, F., Kokordelis, P., Kaczmarek, D., Nischalke, H.D., Strassburg, C.P., Spengler, U. & Nattermann, J.(2016) 'IL28B Genetic Variants Determine the Extent of MonocyteInduced Activation of NK Cells in Hepatitis C' PLoS One. VOL 11

3. Jakubowska, M.A., Ferdek, P.E., Gerasimenko, O.V., Gerasimenko, J.V. & Petersen, O.H.(2016) 'Nitric oxide signals are interlinked with calcium signals in normal pancreatic stellate cells upon oxidative stress and inflammation' Open Biology. VOL 6

The mammalian diffuse stellate cell system comprises retinoid-storing cells capable of remarkable transformations from a quiescent to an activated myofibroblast-like phenotype. Activated pancreatic stellate cells (PSCs) attract attention owing to the pivotal role they play in development of tissue fibrosis in chronic pancreatitis and pancreatic cancer. However, little is known about the actual role of PSCs in the normal pancreas. These enigmatic cells have recently been shown to respond to physiological stimuli in a manner that is markedly different from their neighbouring pancreatic acinar cells (PACs). Here, we demonstrate the capacity of PSCs to generate nitric oxide (NO), a free radical messenger mediating, for example, inflammation and vasodilatation. We show that production of cytosolic NO in PSCs is unambiguously related to cytosolic Ca2+ signals. Only stimuli that evoke Ca2+ signals in the PSCs elicit consequent NO generation. We provide fresh evidence for the striking difference between signalling pathways in PSCs and adjacent PACs, because PSCs, in contrast to PACs, generate substantial Ca2+-mediated and NOS-dependent NO signals. We also show that inhibition of NO generation protects both PSCs and PACs from necrosis. Our results highlight the interplay between Ca2+ and NO signalling pathways in cell–cell communication, and also identify a potential therapeutic target for anti-inflammatory therapies. Less