Acquiring Kupffer Cells in Mice Using a MACS-Based Method

Zhang, C.; Lu, Yaxin; Zhou, Hongliang; Lü, Hongmei; Qian, Xiaofeng; Liu, X.; Wang, X.; Ding, Zhengnian; Zhang, F.; Lü, Ling

doi:10.1016/j.transproceed.2015.01.018

Cited by 6 publications

(4 citation statements)

References 23 publications

(8 reference statements)

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“…Magnetic sorting enriched the CD271 population from around 20% in unsorted SVF, to over 85% in CD271+ AD-MSCs. This is comparable to other groups who have achieved similar high purities with MACS: Zhang et al (2015) produced Kupffer cells of purity 95.7% from mouse livers, after a double MACS sort for F4/80+ and CD11c− cells, and Quirici et al produced a CD271+ population from human AD-MSCs with 88.5% purity by MACS sorting [14,22]. This is despite prior claims that MACS produces significantly lower purities than FACS [23].…”

Section: Discussionsupporting

confidence: 69%

The angiogenic potential of CD271+ human adipose tissue-derived mesenchymal stem cells

et al. 2021

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Background Autologous fat grafting is often a crucial aspect of reconstructive and aesthetic surgeries, yet poor graft retention is a major issue with this technique. Enriching fat grafts with adipose tissue-derived mesenchymal stem cells (AD-MSCs) improves graft survival—however, AD-MSCs represent a heterogeneous population. Selection of subpopulations of AD-MSCs would allow the targeting of specific AD-MSCs that may benefit fat graft survival more than the general AD-MSC population. Methods Human AD-MSCs were selected for the surface marker CD271 using magnetic-activated cell sorting and compared to the CD271 negative phenotype. These subpopulations were analysed for gene expression using Real-Time qPCR and RNA sequencing; surface marker characteristics using immunostaining; ability to form tubules when cultured with endothelial cells; and gene and protein expression of key angiogenic mediators when cultured with ex-vivo adipose tissue. Results Human AD-MSCs with the surface marker CD271 express angiogenic genes at higher levels, and inflammatory genes at lower levels, than the CD271− AD-MSC population. A greater proportion of CD271+ AD-MSCs also possess the typical complement of stem cell surface markers and are more likely to promote effective neoangiogenesis, compared to CD271− AD-MSCs. Conclusion Enriching grafts with the CD271+ AD-MSC subpopulation holds potential for the improvement of reconstructive and aesthetic surgeries involving adipose tissue.

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Section: Discussionsupporting

confidence: 69%

The angiogenic potential of CD271+ human adipose tissue-derived mesenchymal stem cells

et al. 2021

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“…2A). To isolate Kupffer cells from the NPCs, we used a magnetically activated cell sorting (MACS)-based method as described previously (19), with slight modifications. NPCs were incubated with Fc-blocker anti-mouse CD16/32 blocking antibody for 10 min before staining with allophycocyanin (APC)anti-mouse F4/80 antibody.…”

Section: Resultsmentioning

confidence: 99%

Protective Role of Kupffer Cells and Macrophages in Klebsiella pneumoniae-Induced Liver Abscess Disease

2019

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Klebsiella pneumoniae-induced liver abscess (KLA) is emerging as a leading cause of pyogenic liver abscess worldwide. In recent years, the emergence of hypervirulent K. pneumoniae (hvKp) has been strongly associated with KLA. Unlike classical K. pneumoniae, which generally infects the immunocompromised population, hvKp can cause serious and invasive infections in young and healthy individuals. hvKp isolates are often associated with the K1/K2 capsular types and possess hypermucoviscous capsules. KLA is believed to be caused by K. pneumoniae colonizing the gastrointestinal tract of the host and translocating across the intestinal barrier via the hepatic portal vein into the liver to cause liver abscess. We optimized the isolation of the liver-resident macrophages called Kupffer cells in mice and examined their importance in controlling bacterial loads during hvKp infection in healthy mice. Our study reveals the high capability of Kupffer cells to kill hvKp in vitro despite the presence of the bacterial hypermucoviscous capsule, in contrast to other macrophages, which were unable to phagocytose the bacteria efficiently. Depletion of Kupffer cells and macrophages with liposome-encapsulated clodronate (liposomal clodronate) in both an intraperitoneal and an oral mouse infection model resulted in increased bacterial loads in the livers, spleens, and lungs and increased mortality of the infected mice. Thus, Kupffer cells and macrophages are critical for the control of hvKp infection.

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“…, primary KCs and LSECs were isolated from the livers of Swiss Webster mice using a 2‐step collagenase perfusion method. The 2 cell types were enriched using a combination of low‐speed centrifugation, density gradient centrifugation, and Ab‐conjugated magnetic microbeads . For the magnetic separation, anti‐F4/80 microbeads were used to positively select for mouse KCs.…”

Section: Resultsmentioning

confidence: 99%

Sensitivity of Kupffer cells and liver sinusoidal endothelial cells to ricin toxin and ricin toxin–Ab complexes

Mooney

Torres-Vélez

Doering

et al. 2019

Journal of Leukocyte Biology

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Ricin toxin is a plant‐derived, ribosome‐inactivating protein that is rapidly cleared from circulation by Kupffer cells (KCs) and liver sinusoidal endothelial cells (LSECs)—with fatal consequences. Rather than being inactivated, ricin evades normal degradative pathways and kills both KCs and LSECs with remarkable efficiency. Uptake of ricin by these 2 specialized cell types in the liver occurs by 2 parallel routes: a “lactose‐sensitive” pathway mediated by ricin's galactose/N‐acetylgalactosamine‐specific lectin subunit (RTB), and a “mannose‐sensitive” pathway mediated by the mannose receptor (MR; CD206) or other C‐type lectins capable of recognizing the mannose‐side chains displayed on ricin's A (RTA) and B subunits. In this report, we investigated the capacity of a collection of ricin‐specific mouse MAb and camelid single‐domain (VHH) antibodies to protect KCs and LSECs from ricin‐induced killing. In the case of KCs, individual MAbs against RTA or RTB afforded near complete protection against ricin in ex vivo and in vivo challenge studies. In contrast, individual MAbs or VHHs afforded little (<40%) or even no protection to LSECs against ricin‐induced death. Complete protection of LSECs was only achieved with MAb or VHH cocktails, with the most effective mixtures targeting RTA and RTB simultaneously. Although the exact mechanisms of protection of LSECs remain unknown, evidence indicates that the Ab cocktails exert their effects on the mannose‐sensitive uptake pathway without the need for Fcγ receptor involvement. In addition to advancing our understanding of how toxins and small immune complexes are processed by KCs and LSECs, our study has important implications for the development of Ab‐based therapies designed to prevent or treat ricin exposure should the toxin be weaponized.

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Acquiring Kupffer Cells in Mice Using a MACS-Based Method

Cited by 6 publications

References 23 publications

The angiogenic potential of CD271+ human adipose tissue-derived mesenchymal stem cells

The angiogenic potential of CD271+ human adipose tissue-derived mesenchymal stem cells

Protective Role of Kupffer Cells and Macrophages in Klebsiella pneumoniae-Induced Liver Abscess Disease

Sensitivity of Kupffer cells and liver sinusoidal endothelial cells to ricin toxin and ricin toxin–Ab complexes

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