Kupffer Cells

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Kupffer cells are macrophages endogenous to the liver and have the ability to modulate hepatic inflammation and injury associated with various pathophysiologies and toxicities. Pro-inflammatory cytokines released by activated Kupffer cells, such as TNF-α and IL-6, are associated with up-regulation of acute-phase response proteins and suppression of CYP enzymes. For new biological entities, particularly immunomodulators, evaluating the potential for Kupffer cell activation is an emerging concept in preclinical development. Kupffer cells are estimated to comprise approximately 4-8% of total liver cell content and approximately 20% of non-parenchymal cells (Raccanelli and Rehermann, 2006). 

Sekisui XenoTech's Kupffer cell thawing and culturing as well as hepatocyte isolation and plating protocols assure that our fresh plated hepatocyte and Kupffer cell co-cultures contain a representative quantity of liver macrophages.

We examined 19 Sekisui XenoTech freshly plated hepatocyte and Kupffer cell co-cultures for the presence of Kupffer cells. Cells were evaluated for expression of CD68, a general macrophage marker, by immunofluorescent staining after 3 days in culture. Kupffer cells were stained green with an Alexa Fluor® 488-conjugated secondary antibody, while DAPI (4', 6-diamidino-2-phenylindole) was used as a nuclear counterstain. Kupffer cell content was quantified from 3 images taken from separate wells in a 24-well plate.

Sekisui XenoTech's fresh plated hepatocyte and Kupffer cell co-cultures contain 0.6% to 24.1% Kupffer cells with a median value of 2.6% and average of 4.7%. The cytokine response of isolated Kupffer cells to stimulation with bacterial endotoxin (LPS) in vitro has been characterized by Sekisui XenoTech's scientists (Lambert et al., 2013).

The images below illustrate typical composition of the hepatocyte and Kupffer cell co-cultures of four liver donors:

Kuppfer_Cells.jpg
(A) 43 year old Caucasian female, 9.3% Kupffer cells, (B) 45 year old Caucasian male, 2.2% Kupffer cells, (C) 7 year old Caucasian male, 3.6% Kupffer cells, (D) 53 year old Hispanic female, 5.4% Kupffer cells. The images shown, selected to emphasize Kupffer cell morphology, are only a part of the field that was used for quantification of the macrophages.
 
XenoTech offers Kupffer cells to aide in the assessment of cytokine release by biologics and small molecule drugs. Our Kupffer cells have been characterized for the following:
 
  • Cell viability and cell yield (by Trypan blue staining)
  • Functionality:  Activation by bacterial lipopolysaccharide (LPS) measured by induction of Tumor Necrosis Factor-alpha (TNF-a) and Interleukin-6 (IL-6) mRNA and protein levels
  • Purity:  Expression of macrophage marker CD68 (by Immuno-fluorescent microscopy)

  Buy cell and tissue based products
Product Description Volume AMY
HK1000.H15 Cryopreserved male human Kupffer Cells 1mL 1 million
HK1500.H15 Cryopreserved female human Kupffer Cells 1mL 1 million
K8700 OptiThaw Kupffer Cell Thaw/Culture Media 100mL
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