Cryo-imaging of Stem Cell Biodistribution in Mouse Model of Graft-Versus-Host-Disease

Wuttisarnwattana, Patiwet; Eid, Saada; Gargesha, Madhusudhana; Cooke, Kenneth R.; Wilson, David L.

doi:10.1007/s10439-020-02487-z

Cited by 14 publications

(18 citation statements)

References 28 publications

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“…To address this unmet need, we developed a novel lymphocyte proliferation assessment protocol based on cryo-imaging. Cryo-imaging 35 – 38 is a fully-automated, whole mouse, section-and-image system, which provides 3D, tiled, microscopic, anatomical brightfield and molecular fluorescent images over vast volumes. It provides single cell detection anywhere in the mouse and determines cell densities far below what can be observed with any other imaging technologies such as MRI, CT, PET, SPECT and BLI.…”

Section: Introductionmentioning

confidence: 99%

Assessment of the therapeutic role of mesenchymal stromal cells in a mouse model of graft-versus-host disease using cryo-imaging

Wuttisarnwattana

Eid

Wilson

et al. 2023

Sci Rep

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Insights regarding the biodistribution and homing of mesenchymal stromal cells (MSCs), as well as their interaction with alloreactive T-cells are critical for understanding how MSCs can regulate graft-versus-host disease (GVHD) following allogeneic (allo) bone marrow transplantation (BMT). We developed novel assays based on 3D, microscopic, cryo-imaging of whole-mouse-sized volumes to assess the therapeutic potential of human MSCs using an established mouse GVHD model. Following infusion, we quantitatively tracked fluorescently labeled, donor-derived, T-cells and third party MSCs in BMT recipients using multispectral cryo-imaging. Specific MSC homing sites were identified in the marginal zones in the spleen and the lymph nodes, where we believe MSC immunomodulation takes place. The number of MSCs found in spleen of the allo BMT recipients was about 200% more than that observed in the syngeneic group. To more carefully define the effects MSCs had on T cell activation and expansion, we developed novel T-cell proliferation assays including secondary lymphoid organ (SLO) enlargement and Carboxyfluoescein succinimidyl ester (CFSE) dilution. As anticipated, significant SLO volume enlargement and CFSE dilution was observed in allo but not syn BMT recipients due to rapid proliferation and expansion of labeled T-cells. MSC treatment markedly attenuated CFSE dilution and volume enlargement of SLO. These assays confirm evidence of potent, in vivo, immunomodulatory properties of MSC following allo BMT. Our innovative platform includes novel methods for tracking cells of interest as well as assessing therapeutic function of MSCs during GVHD induction. Our results support the use of MSCs treatment or prevention of GVHD and illuminate the wider adoption of MSCs as a standard medicinal cell therapy.

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Section: Introductionmentioning

confidence: 99%

Assessment of the therapeutic role of mesenchymal stromal cells in a mouse model of graft-versus-host disease using cryo-imaging

Wuttisarnwattana

Eid

Wilson

et al. 2023

Sci Rep

Self Cite

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“…Furthermore, a full characterization of the infused MSCs was carried out, but neither in vitro functional assay to determine their immunomodulatory potency nor biodistribution analysis was performed. However, previous studies have shown that after intravenous infusion, cells were accumulated in lung, spleen, liver, and bone marrow 37 …”

Section: Discussionmentioning

confidence: 96%

“…However, previous studies have shown that after intravenous infusion, cells were accumulated in lung, spleen, liver, and bone marrow. 37 On the other hand, a higher number of patients and a control group, planned in the second phase of the trial, would have allowed us to better assess the fluctuations that some patients presented in several immunological parameters. Furthermore, the analyses are based on PBMC samples, which may not always reliably reflect tissuerelated processes during HIV infection.…”

Section: Discussionmentioning

confidence: 99%

Mesenchymal stromal cells in human immunodeficiency virus-infected patients with discordant immune response: Early results of a phase I/II clinical trial

Trujillo‐Rodríguez

Viciana

Rivas‐Jeremías

et al. 2020

Stem Cells Translational Medicine

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Between 15% and 30% of HIV-infected subjects fail to increase their CD4 + T-cell counts despite continuous viral suppression (immunological nonresponders [INRs]). These subjects have a higher morbidity and mortality rate, but there are no effective treatments to reverse this situation so far. This study used data from an interrupted phase I/II clinical trial to evaluate safety and immune recovery after INRs were given four infusions, at baseline and at weeks 4, 8, and 20, with human allogeneic mesenchymal stromal cells from adipose tissue (Ad-MSCs). Based on the study design, the first 5 out of 15 INRs recruited received unblinded Ad-MSC infusions. They had a median CD4 + nadir count of 16/μL (range, 2-180) and CD4 + count of 253 cells per microliter (171-412) at baseline after 109 (54-237) months on antiretroviral treatment and 69 (52-91) months of continuous undetectable plasma HIV-RNA. After a year of follow-up, an independent committee recommended the suspension of the study because no increase of CD4 + T-cell counts or CD4 + /CD8 + ratios was observed. There were also no significant changes in the phenotype of different immunological lymphocyte subsets, percentages of natural killer cells, regulatory T cells, and dendritic cells, the inflammatory parameters analyzed, and cellular associated HIV-DNA in peripheral blood mononuclear cells. Furthermore, three subjects suffered venous thrombosis events directly related to the Ad-MSC infusions in the arms where the infusions were performed. Although the current study is based on a small sample of participants, the findings suggest that allogeneic Ad-MSC infusions are not effective to improve immune recovery in INR patients or to reduce immune activation or inflammation. ClinicalTrials.gov identifier: NCT0229004.

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“…Implants harvested at postoperative days 1, 3, and 7 were subjected to fluorescence-activated cell sorting (FACs), RT-PCR, and western blot. At 7 days, wild GFP + mBMSCs or cells intervened by shRNA were injected (1 × 10 6 /mouse) via tail vein every 2 days (Supplemental Figure 1 ) [ 16 , 17 ].…”

Section: Methodsmentioning

confidence: 99%

Endothelial Cells Promote Migration of Mesenchymal Stem Cells via PDGF-BB/PDGFRβ-Src-Akt in the Context of Inflammatory Microenvironment upon Bone Defect

Hou

Zhou

et al. 2022

Stem Cells International

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Homing of mesenchymal stem cells (MSCs) to the defect site is indispensable for bone repair. Local endothelial cells (ECs) can recruit MSCs; however, the mechanism remains unclear, especially in the context of the inflammatory microenvironment. This study was aimed to investigate the role of ECs in MSCs migration during the inflammatory phase of bone repair. The inflammatory microenvironment was mimicked in vitro via adding a cytokine set (IL-1β, IL-6, and TNF-α) to the culture medium of ECs. The production of PDGF-BB from ECs was measured by ELISA. Transwell and wound healing assays were employed to assess MSCs migration toward ECs and evaluate the implication of PDGF-BB/PDGFRβ. A series of shRNA and pathway inhibitors were used to screen signal molecules downstream of PDGF-BB/PDGFRβ. Then, mouse models of femoral defects were fabricated and DBM scaffolds were implanted. GFP+ MSCs were injected via tail vein, and the relevance of PDGF-BB/PDGFRβ, as well as screened signal molecules, in cell homing was further verified during the early phase of bone repair. In the mimicked inflammatory microenvironment, MSCs migration toward ECs was significantly promoted, which could be abrogated by pdgfrb knockout in MSCs. Inhibition of Src or Akt led to negative effects analogous to pdgfrb knockout. Blockade of JNK, MEK, and p38 MAPK had no impact. Meanwhile, the secretion of PDGF-BB from ECs was evidently motivated by the inflammatory microenvironment. Adding recombinant PDGF-BB protein to the culture medium of ECs phenocopied the inflammatory microenvironment with regard to attracting MSCs, which was abolished by pdgfb, src, or akt in MSCs. Moreover, pdgfb knockout suppressed the expression and phosphorylation of Src and Akt in migrating MSCs. Src knockout impaired Akt expression but not vice versa. In vivo, reduced infiltration of CD31+ ECs was correlated with diminished PDGF-BB in local defect sites, and silencing pdgfb, src, or akt in MSCs markedly hampered cell homing. Together, these findings suggest that in the inflammatory microenvironment, MSCs migrate toward ECs via PDGF-BB/PDGFRβ and the downstream Src-Akt signal pathway.

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Cryo-imaging of Stem Cell Biodistribution in Mouse Model of Graft-Versus-Host-Disease

Cited by 14 publications

References 28 publications

Assessment of the therapeutic role of mesenchymal stromal cells in a mouse model of graft-versus-host disease using cryo-imaging

Assessment of the therapeutic role of mesenchymal stromal cells in a mouse model of graft-versus-host disease using cryo-imaging

Mesenchymal stromal cells in human immunodeficiency virus-infected patients with discordant immune response: Early results of a phase I/II clinical trial

Endothelial Cells Promote Migration of Mesenchymal Stem Cells via PDGF-BB/PDGFRβ-Src-Akt in the Context of Inflammatory Microenvironment upon Bone Defect

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