Lysosomal Enzyme Content of Kupffer and Endothelial Liver Cells Isolated from Germfree and Clean Conventional Rats

Knook, D.L.; Barkway, Chris; Sleyster, E.Ch.

doi:10.1128/iai.33.2.620-622.1981

Cited by 17 publications

(19 citation statements)

References 14 publications

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“…The perfusion step could be followed by incubation in the same solution that was associated [ 19 , 20 ] or not [ 18 , 21 , 22 ] with DNase for as much as 30 minutes [ 18 , 22 ]. Another study described the use of Pronase for liver digestion [ 39 ].…”

Section: Resultsmentioning

confidence: 99%

“…In rats, another method for non-parenchymal cell isolation described in the 1980s consisted of liver perfusion and digestion by Pronase to destroy parenchymal cells, in association with centrifugation in a one-step gradient of Metrizamide [ 22 , 39 ] or a four-step gradient of Stractan [ 30 ].…”

Section: Resultsmentioning

confidence: 99%

“…LSEC were purified from the cell fraction devoid of parenchymal cells by centrifugal elutriation in 37.50% of the publications (6/16) [ 12 , 18 , 22 , 31 , 35 , 39 ], by selective adherence in 43.75% (7/16) [ 19 – 21 , 30 , 33 , 34 , 36 ] and by SE-1-based MACS in 6.25% (1/16) [ 36 ]. No purification step following gradient centrifugation was reported in 2 publications [ 37 , 38 ].…”

Section: Resultsmentioning

confidence: 99%

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Methods for Isolation and Purification of Murine Liver Sinusoidal Endothelial Cells: A Systematic Review

et al. 2016

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To study the biological functions of liver sinusoidal endothelial cells (LSEC) and to identify their interplay with blood or liver cells, techniques allowing for the isolation and purification of LSEC have been developed over the last decades. The objective of the present review is to summarize and to compare the efficiency of existing methods for isolating murine LSEC. Toward this end, the MEDLINE database was searched for all original articles describing LSEC isolation from rat and mouse livers. Out of the 489 publications identified, 23 reported the main steps and outcomes of the procedure and were included in our review. Here, we report and analyse the technical details of the essential steps of the techniques used for LSEC isolation. The correlations between the prevalence of some steps and the efficiency of LSEC isolation were also identified. We found that centrifugal elutriation, selective adherence and, more recently, magnetic-activated cell sorting were used for LSEC purification. Centrifugal elutriation procured high yields of pure LSEC (for rats 30–141.9 million cells for 85–98% purities; for mice 9–9.25 million cells for >95% purities), but the use of this method remained limited due to its high technical requirements. Selective adherence showed inconsistent results in terms of cell yields and purities in rats (5–100 million cells for 73.7–95% purities). In contrast, magnetic-activated cell sorting allowed for the isolation of highly pure LSEC, but overall lower cell yields were reported (for rats 10.7 million cells with 97.6% purity; for mice 0.5–9 million cells with 90–98% purities). Notably, the controversies regarding the accuracy of several phenotypic markers for LSEC should be considered and their use for both magnetic sorting and characterization remain doubtful. It appears that more effort is needed to refine and standardize the procedure for LSEC isolation, with a focus on the identification of specific antigens. Such a procedure is required to identify the molecular mechanisms regulating the function of LSEC and to improve our understanding of their role in complex cellular processes in the liver.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Methods for Isolation and Purification of Murine Liver Sinusoidal Endothelial Cells: A Systematic Review

et al. 2016

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“…Patients. The sera of patients with acute hepatitis A were collected at the time of admission to the hospital (To) and at 6 weeks (T1), 3,6, and 12 months later. In each case, a hepatitis B was serologically excluded (negative for hepatitis B surface antigen and IgM antihepatitis B core).…”

Section: Methodsmentioning

confidence: 99%

Increase in immunoglobulin M antibodies against gut bacteria during acute hepatitis A

Miller¹,

Legler²,

Thomssen³

1983

Infect Immun

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The marked increase in the total serum immunoglobulin M (IgM) is a characteristic feature of acute hepatitis A. To study the nature of this IgM, we assayed serial titers of IgM antibodies against various antigens during and after acute hepatitis A. The antibodies against blood group antigen remained unchanged throughout the observation period. Thus, the production or metabolism of IgM was not nonspecifically altered. The IgM antibody against hepatitis A antigen decreased and finally disappeared during convalescence as expected. However, its time course did not correlate quantitatively with the concentration of the total serum IgM. In contrast, IgM antibodies against gut bacteria Bacteroides fragilis and Streptococcus faecalis were considerably elevated in all patients at the onset of the disease, and they normalized similarly to the total IgM during convalescence. IgM antibodies against Escherichia coli were elevated only in some of the patients. The data suggest that the amount of IgM antibodies against gut bacteria contributes significantly to the increase in the total serum IgM in acute hepatitis A.

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“…It is not clear whether macrophage functions from every aspect are not fully matured or expressed in germfree mice. Many reports indicated a poor expression of some macrophage functions in germfree animals (2,5,11), although there are some showing no difference between germfree and conventional mice (7,20). Bacterial flora consist of various species of bacteria, and gram-negative bacteria especially reside as intestinal microflora.…”

Section: Discussionmentioning

confidence: 99%

Ontogeny of macrophage function to release superoxide anion in conventional and germfree mice

Mitsuyama¹,

Ohara²,

Amako³

et al. 1986

Infect Immun

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Lysosomal Enzyme Content of Kupffer and Endothelial Liver Cells Isolated from Germfree and Clean Conventional Rats

Abstract: Rats kept under germfree conditions showed lower specific lysosomal enzyme activities in liver endothelial cells, but not in Kupffer cells.

Cited by 17 publications

References 14 publications

Methods for Isolation and Purification of Murine Liver Sinusoidal Endothelial Cells: A Systematic Review

Methods for Isolation and Purification of Murine Liver Sinusoidal Endothelial Cells: A Systematic Review

Increase in immunoglobulin M antibodies against gut bacteria during acute hepatitis A

Ontogeny of macrophage function to release superoxide anion in conventional and germfree mice

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