geoscience.net logo
+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Development of a cytokine-producing immortalized murine Kupffer cell line


, : Development of a cytokine-producing immortalized murine Kupffer cell line. Cytokine 70(2): 165-172

Kupffer cells (KC) play a critical role in both liver physiology and the pathogenesis of various liver diseases. Isolated primary KC have a limited lifespan in culture, and due to the relatively low number obtained, limit their study in vitro. Here, a cytokine-producing immortalized KC (ImKC) line was established from transgenic mice that express the thermolabile mutant tsA58 of the Simian virus 40 large T antigen under the control of the H-2k(b) promoter. Primary KC were obtained using a three step procedure: liver perfusion, centrifugal elutriation, and sorting for F4/80⁺ cells. ImKC were identified within the small-intermediate population of KC that maintained stable expression of F4/80, and the surface antigens CD11b, CD14 and TLR4. ImKC grow at IFNγ-independent manner at 37°C and exhibited a doubling time of ∼24 h when cultured in RPMI 1640 with 5% FBS. Our observations indicate that both activation of telomerase and expression of P53 are markedly increased, suggesting that enhanced telomerase activity and P53 expression may contribute to the immortalization of this cell population. ImKC cells maintained a high capacity to phagocytose FITC-latex beads, and bind/phagocytose erythrocytes. In addition, similar to primary KC, ImKC responded to stimulation with lipopolysaccharide (LPS: 0.1-1μg/ml) by upregulating mRNA levels of TNFα (23-fold), IL-6 (28-fold), and IL-1β (1459-fold), as measured by qRT-PCR. Protein levels of TNFα and IL-6 were also increased, 10-fold and 12-fold, respectively. Reactive oxygen species (ROS) and nitric oxide (NO) production were significantly enhanced in ImKC following an LPS challenge. Furthermore, LPS elicited a marked increase in mitogen activated protein kinase (MAPK) phospho-(ERK1/2, JNK) and NF-κB p50 with decreased IκBα in ImKC, as assessed by Western blot. Collectively, these results demonstrate that the ImKC line retains critical characteristics of primary KC, and thus provides a useful tool to assess the role of KC in liver injury and chronic diseases.

US$19.90

PMID: 25138015

DOI: 10.1016/j.cyto.2014.07.251


Other references

Stewart T.P.; Hui S.W.; Portes A.; Papahadjopoulos D., 1979: Complex phase mixing in di palmitoyl phosphatidyl choline bovine brain phosphatidyl serine system. Biophysical Journal 25(2 PART 2): 174A

Reuss, F.U.; Frankel, W.N.; Moriwaki, K.; Shiroishi, T.; Coffin, J.M., 1996: Genetics of intracisternal-A-particle-related envelope-encoding proviral elements in mice. Intracisternal-A-particle-related envelope-encoding (IAPE) proviral elements in the mouse genome encode and express an envelope-like protein that may allow transmission of IAPEs as infectious agents. To test IAPE mobility and potential transmissio...

Moraes, M.P.; Weiblen, R.; Rebelatto, M.C.; Silva, A.M. da, 2000: Relationship between passive immunity and morbidity and weight gain in dairy cattle. Serum samples were collected from 87 calves from a dairy herd in Rio Grande do Sul, Brazil. Serum immunoglobulin (Ig) levels were measured by the zinc sulfate turbidity test at 24 hours old. The mean serum Ig level was 11.40 g/litre. 14 out of 87...

Chuanhao Chen; B.X.e; Yuan Ren; Chaofei Wu; Chaohai Wei, 2000: The mechanisms of affecting factors in treating wastewater by Fenton reagent. The mechanisms of affecting factors in treating toxic organic wastewater such as wood-glue wastewater by Fenton reagent were preliminarily investigated. The optimal operating conditions determined by orthogonal experiments are as following: (H2O2)...

Rogovskaia, T.I., 1951: Effect of trinitrotoluol on microorganisms and biochemical processes in autopurification of water. Mikrobiologiia 20(3): 265-272

Rembold, C.M.; Murphy, R.A., 1989: Histamine concentration and Ca2+ mobilization in arterial smooth muscle. Smooth muscle contraction is dependent on Ca2+ entry from the extracellular space or release from intracellular stores. The sensitivity of these Ca2+ sources to agonist concentration was evaluated by measuring myoplasmic [Ca2+] (as estimated by ae...

Plona T.J.; Tsang L., 1978: Characterization of the average microscopic dimension in granular media using ultrasonic pulses; theory and experiments. Abstracts - Society of Exploration Geophysicists International Meeting 48): 38-39

Robinson, D.S., 1965: The clearing factor lipase activity of adipose tissue. Handbook Physiol 5: 295-299

Hodgson, L.; Pertz, O.; Hahn, K.M., 2007: Design and optimization of genetically encoded fluorescent biosensors: GTPase biosensors. This chapter details the design and optimization of biosensors based on a design used successfully to study nucleotide loading of small GTPase proteins in living cells. This design can be generalized to study many other protein activities, using a...

Minter, D.W., 1988: Ploioderma cedri.. A description is provided for Ploioderma cedri. Information is included on the disease caused by the organism, its transmission, geographical distribution, and hosts. HOSTS: Cedrus deodara. DISEASE: This fungus is a pathogen of Cedrus deodara, cau...