Isolation and primary culture of rat Kupffer cells

Olynyk, John K.; Clarke, Sharon L

doi:10.1111/j.1440-1746.1998.tb00746.x

Cited by 34 publications

(29 citation statements)

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“…The cells were finally obtained after centrifugation (4 C, 100g, 5 min) and were seeded into 24-well plates in 1 ml of Dulbecco's Modified Eagle's Medium with 10% fetal bovine serum (FBS). KCs were isolated from mice under pentobarbitone anesthesia Olynyk et al, 1998). In brief, the rat portal vein was cannulated and perfused with Hank's buffered salt solution for 10 min, the liver was excised, and the perfusate was discarded.…”

Section: In Vivo Transfection Studiesmentioning

confidence: 99%

Dual ligands modified double targeted nano-system for liver targeted gene delivery

Jing

Liu

et al. 2013

Pharmaceutical Biology

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Context: It is now well established that the surface of nanocarriers with specific ligands defines a new biological identity, which assist in targeting and internalization of the nanocarriers to specific cell populations, such as cancers and disease organs. Objective: The aim of this study is to develop systemically administrable dual ligands modified nano-system which could both target cancer cells and macrophages in the liver. Methods: Transferrin (Tf) and mannan (M) were linked onto polyethylene glycol-phosphatidylethanolamine (PEG-PE) and PE separately to get transferrin-PEG-PE (T-PEG-PE) and mannan-PE

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Section: In Vivo Transfection Studiesmentioning

confidence: 99%

Dual ligands modified double targeted nano-system for liver targeted gene delivery

Jing

Liu

et al. 2013

Pharmaceutical Biology

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“…Kupffer cells were har vested from the liver using isolation buffers according to a previously described procedure [13,14] . The livers were perfused in situ with Ca 2+ -free minimum essential medium (Sigma, St. Louis, MI, USA), followed by 0.3% pronase (Roche Diagnostics Co., Indianapolis, IN, USA) and 0.05% collagenase type Ⅳ (Wako, Osaka, Japan) in Dulbecco's modified Eagle's medium/F-12 (Sigma) at a rate of 10 mL/min through the portal vein.…”

Section: Isolation and Culture Of Kupffer Cellsmentioning

confidence: 99%

Immunotherapy for nonalcoholic steatohepatitis using themultiple cytokine production modulator Y-40138

Tsujimoto¹,

Kawaratani²,

Kitazawa³

et al. 2009

WJG

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AIM:To investigate the possible use of the multiple cytokine production modulator, Y-40138, as a novel immunotherapy in the rat nonalcoholic steatohepatitis (NASH) model. METHODS:We allocated 6-wk-old male F344 rats to choline-supplemented, L-amino acid-defined (CSAA) diet (control group), CSAA diet + Y-40138 (control + Y-40138 group), choline-deficient, L-amino acid-defined (CDAA) diet (NASH group), or CDAA diet + Y-40138 (NASH + Y-40138 group). In each group, we measured the plasma alanine aminotransferase (ALT) levels, and the plasma and liver levels of tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), and interleukin-10 (IL-10). Tissue specimens of phosphate buffered saline-perfused liver were subjected to hematoxylin and eosin staining, Azan staining, Sirius red staining, and immunohistochemical staining (for Kupffer cells and TNF-α). We then extracted Kupffer cells from the collagenase-perfused livers using the Percoll gradient centrifugation method, and measured the TNF-α levels in the supernatant (in vitro TNF-α production by Kupffer cells) using an enzyme-linked immunosorbent assay kit. RESULTS:In comparison to the NASH group, serum ALT elevation was mild, production of serum and liver TNF-α and IFN-γ was inhibited, and IL-10 production was increased in the NASH + Y-40138 group. Amelioration of liver histology was also noted in the NASH + Y-40138 group. Kupffer cell immunohistochemical staining revealed no differences between groups, whereas TNF-α immunohistochemical staining showed fewer stained cells in the NASH + Y-40138 group than in the NASH group. The TNF-α levels in the in-vitro Kupffer cell culture supernatant were lower in the NASH + Y-40138 group than in the NASH group. CONCLUSION:Administration of Y-40138 to NASH model rats reduced hepatic inflammation and suppressed fibrosis. These results indicate that the multiple cytokine production modulator, Y-40138, is promising as a novel treatment for NASH.

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“…Kupffer cells were isolated from the control and CDAAfed rat livers according to a previously described procedure [10,11] . In brief, the liver was perfused through the portal vein with 100 mL of PBS containing 0.5 mmol/L EGTA (Research Organics, Cleveland, OH, USA) solution.…”

Section: Kupffer Cell Isolation and Culturementioning

confidence: 99%

Innate immune reactivity of the liver in rats fed a cholinedeficient L-amino-acid-defined diet

Kawaratani¹,

Tsujimoto²,

Kitazawa³

et al. 2008

WJG

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AIM:To investigate the innate immune reactivity of tumor necrosis factor-alpha (TNF-α), Toll-like receptor 4 (TLR4), and CD14 in the liver of non-alcoholic steatohepatitis (NASH) model rats. METHODS: Male F344 rats were fed a cholinedeficient L-amino-acid-defined (CDAA) diet. The rats were killed after 4 or 8 wk of the diet, and their livers were removed for immunohistochemical investigation and RNA extraction. The liver specimens were immunostained for TNF-α, TLR4, and CD14. The gene expressions of TNF-α , TLR4 , and CD14 were determined by reverse-transcriptase polymerase chain reaction (RT-PCR). Kupffer cells were isolated from the liver by Percoll gradient centrifugation, and were then cultured to measure TNF-α production. RESULTS: The serum and liver levels of TNF-α in the CDAA-fed rats increased significantly as compared with the control group, as did the immunohistochemical values and gene expressions of TNF-α, TLR4, and CD14 with the progression of steatohepatitis. TNF-α production from the isolated Kupffer cells of the CDAAfed rats was elevated by lipopolysaccharide stimulation. CONCLUSION: The expressions of TNF-α, TLR4, and CD14 increased in the NASH model, suggesting that TLR4 and CD14-mediated endotoxin liver damage may also occur in NASH.

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Isolation and primary culture of rat Kupffer cells

Cited by 34 publications

References 0 publications

Dual ligands modified double targeted nano-system for liver targeted gene delivery

Dual ligands modified double targeted nano-system for liver targeted gene delivery

Immunotherapy for nonalcoholic steatohepatitis using themultiple cytokine production modulator Y-40138

Innate immune reactivity of the liver in rats fed a cholinedeficient L-amino-acid-defined diet

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