A New Method to Isolate and Culture Rat Kupffer Cells

Zeng, Weiqun; Zhang, Ji-qin; Li, Yue; Yang, Kang; Yupei, Chen; Liu, Zuojin

doi:10.1371/journal.pone.0070832

Cited by 45 publications

(39 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Liver macrophages were isolated from mouse livers as reported previously (46) with some modifications. Before collection of the tissue, mice were culled and bled by cutting the inferior vena cava.…”

Section: Methodsmentioning

confidence: 99%

Local amplifiers of IL-4Rα–mediated macrophage activation promote repair in lung and liver

Minutti

Jackson-Jones

García-Fojeda

et al. 2017

Science

175

170

View full text Add to dashboard Cite

The type 2 immune response controls helminth infection and maintains tissue homeostasis but can lead to allergy and fibrosis when improperly controlled. We reveal the existence of local tissue-specific enhancers of type 2 mediated-macrophage activation. In the lung, surfactant protein A (SP-A) enhanced IL-4-dependent proliferation and activation of alveolar macrophages, accelerating parasite clearance and reducing pulmonary injury following infection with a lung-migrating helminth. In the peritoneal cavity and liver, C1q enhancement of type 2 macrophage activation was required for liver repair following bacterial infection, but resulted in tissue fibrosis following peritoneal dialysis. Both SP-A and C1q generated their effects on macrophages via the unconventional myosin18A that acts as a cell surface receptor. We conclude that the structurally related defense collagens SP-A and C1q are tissue-specific factors that act through myosin18A to license local type 2 responses with consequences for parasite control, tissue repair, and fibrosis.

show abstract

Section: Methodsmentioning

confidence: 99%

Local amplifiers of IL-4Rα–mediated macrophage activation promote repair in lung and liver

Minutti

Jackson-Jones

García-Fojeda

et al. 2017

Science

175

170

View full text Add to dashboard Cite

show abstract

“…[24][25][26] After iv administration of sodium heparin (100-200 units), the livers were exsanguinated in situ by portal vein perfusion with Ca 2+ -free Hanks' solution (pH 7.4), at a constant rate of 10-20 mL/min, and subsequently incubated with fresh collagenase buffer at 37°C for 5 min. The liver was then excised and transferred into precooled PBS at 4°C; then the liver membrane was cut, the liver was shaken gently, and the liver tissue was homogenized using a glass needle.…”

Section: Kc Isolationmentioning

confidence: 99%

Effect of inhibitors of endocytosis and NF-kB signal pathway on folate-conjugated nanoparticle endocytosis by rat Kupffer cells

Tang¹,

Chen²,

Jia³

et al. 2017

IJN

View full text Add to dashboard Cite

The regular accumulation of nanoparticles in the liver makes them hepatotoxic and decreases the circulation time, thus reducing their therapeutic effect. Resolving this problem will be significant in improving bioavailability and reducing side effects. In this study, we reduced the phagocytosis of epirubicin (EPI)-loaded folic acid-conjugated pullulan acetate (FPA/EPI) nanoparticles by Kupffer cells (KCs) through internalization and nuclear factor kappa B (NF-kB) signal pathway inhibitors, thus allowing development of FPA/EPI nanoparticles as a nanodrug delivery system (NDDS) based on our previous study. FPA/EPI nanoparticles were prepared by the dialysis method. Rat KCs were preincubated with the following individual or compound inhibitors: chlorpromazine (CPZ), nystatin (NY), colchicine (Col), amiloride (AMR), and pyrrolidine dithiocarbamate (PDTC). Dose- and time-dependent cellular uptake effects of inhibitors on FPA/EPI nanoparticles were determined through fluorometry. The cytokine levels of tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β), and IL-6 were tested in culture supernatants by bead-based multiplex flow cytometry. The uptake study demonstrated that inhibitors had an obvious inhibitory effect ( P <0.05 or P <0.01), with NY, AMR and Col all showing time-dependent inhibitory effects. PDTC + NY had the strongest inhibitory effect, with an uptake rate of 14.62%. The levels of the three proinflammatory cytokines were changed significantly by the compound inhibitors. TNF-α was significantly inhibited ( P <0.05 or P <0.01), but IL-1β and IL-6 showed smaller decreases. These results suggested that clathrin- and caveolae-mediated endocytosis were the main routes via which nanoparticles entered KCs and that the NF-kB signal pathway was very important too. In summary, multiple mechanisms, including clathrin- and caveolae-mediated endocytosis, contribute to cytokine production in macrophages following exposure to folic acid-conjugated pullulan acetate nanoparticles. Thus, the endocytosis inhibition strategy has great potential for improving therapy and reducing toxicity of an NDDS in the treatment of cancer.

show abstract

“…KCs were harvested by collagenase digestion, differential centrifugation, and selective adherence as described previously [17]. The viability of the isolated cells was determined by 0.1 % trypan blue (Sigma, USA) exclusion and routinely exceeded 90 %.…”

Section: Isolation and Culture Of Kcsmentioning

confidence: 99%

Mesenchymal stromal cell-dependent reprogramming of Kupffer cells is mediated by TNF-α and PGE2 and is crucial for liver transplant tolerance

et al. 2015

Self Cite

View full text Add to dashboard Cite

The role of mesenchymal stromal cells (MSCs) in the modulation of liver transplant tolerance has attracted significant interest. However, the interaction between MSCs and Kupffer cells (KCs) has received little attention, and the effect of this interaction on liver transplant tolerance remains unclear. KCs were cultured in the presence and absence of MSCs. After 24 h, cells were treated with lipopolysaccharide (LPS), after which the production of cytokines and the expression of surface antigens were measured for cell function identification. Moreover, the effects of the KCs and the prostaglandin E2 (PGE2) levels produced by the MSCs were determined using an experimental rat liver transplantation model. Blood and liver samples were collected at three time points after transplantation for further analysis. After LPS treatment, when compared with the KC single cultures, the expression of pro-inflammatory cytokines (IL-1β, IL-6, MHC-II, CD40, CD80, and CD86) in the coculture system was down-regulated, whereas the expression of anti-inflammatory cytokines (TGF-β, IL-4, PGE2, and IL-10) was markedly increased. These data indicate that MSCs can reprogram the phenotype of KCs. However, KCs treated with miR/TNF-α (tumor necrosis factor) plasmid prior to coculture to inhibit the production of TNF-α resulted in an inhibition of the reprogramming effect of MSCs. Moreover, overexpression of PGE2 in MSCs increased the effect of MSCs on KC reprogramming. After rat liver transplantation, allograft recipients that received MSCs showed better allograft tolerance when compared with rats in which KC function was inhibited. Furthermore, rats treated with MSCs overexpressing PGE2 demonstrated the best liver tolerance of all of the groups tested. MSCs reprogram the phenotype of KCs through TNF-α and PGE2, and this process is crucial for the immunomodulatory function of MSCs in liver transplantation.

show abstract

A New Method to Isolate and Culture Rat Kupffer Cells

Cited by 45 publications

References 33 publications

Local amplifiers of IL-4Rα–mediated macrophage activation promote repair in lung and liver

Local amplifiers of IL-4Rα–mediated macrophage activation promote repair in lung and liver

Effect of inhibitors of endocytosis and NF-kB signal pathway on folate-conjugated nanoparticle endocytosis by rat Kupffer cells

Mesenchymal stromal cell-dependent reprogramming of Kupffer cells is mediated by TNF-α and PGE2 and is crucial for liver transplant tolerance

Contact Info

Product

Resources

About