Flow cytometry enumeration and characterization of stem cell lineages in the liver of patients with hepatic tumors

Abstract Background Severe, steroid-resistant asthma (SSRA) is a serious clinical problem in asthma management. Affected patients have severe clinical symptoms, worsened quality of life, and do not respond to steroid, a mainstay steroid treatment of asthma. Thus, effective therapies are urgently needed. Exosomes derived from mesenchymal stem cell (MSC-Exo) has become attractive candidates for the lung inflammatory diseases through its immunomodulatory effects. In this study, we explored the therapeutic effects of MSC-Exo in SSRA and identified the therapeutic mechanism of MSC-Exo. Method Exosomes from human umbilical cord mesenchymal stem cell (hUCMSC) were isolated and characterized by transmission electron microscopy, nanoparticle tracking analysis and flow cytometry analysis. Effects of MSC-Exo on airway hyper responsiveness (AHR), inflammation, histopathology, and macrophage polarization in SSRA in mice were evaluated. Systematic depletion of macrophages determined the role of macrophages in the therapeutic effect of SSRA in mice. LPS-stimulated RAW 264.7 cell model was constructed to determine the underlying mechanism of MSC-Exo on macrophage polarization. qRT-PCR, Western blotting, immunofluorescence, and flow cytometry were performed to evaluate the expression of M1 or M2 markers. Tandem mass tags (TMT)-labeled quantitative proteomics were applied to explore the central protein during the regulation effect of MSC-Exo on macrophage polarization. Knockdown and overexpression of TRAF1 were used to further clarify the role of the central protein on macrophage polarization. Result We successfully isolated and characterized exosomes from hUCMSCs. We verified that the intratracheal administration of MSC-Exo reversed AHR, histopathology changes, and inflammation in SSRA mice. Systematic depletion of macrophages weakened the therapeutic effect of MSC-Exo. We found that MSC-Exo treatment inhibited M1 polarization and promoted M2 polarization in LPS-stimulated RAW 264.7 cells. Subsequently, tumor necrosis factor receptor-associated factor 1 (TRAF1) was determined as the central protein which may be closely related to the regulation of macrophage polarization from TMT-labeled quantitative proteomics analysis. Knockdown and overexpression of TRAF1 demonstrated that the effect of MSC-Exo treatment on macrophage polarization, NF-κB and PI3K/AKT signaling was dependent on TRAF1. Conclusion MSC-Exo can ameliorate SSRA by moderating inflammation, which is achieved by reshaping macrophage polarization via inhibition of TRAF1.

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Selection and characterization of F9 teratocarcinoma stem cell mutants with altered responses to retinoic acid.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)91100-2 ◽

1984 ◽

Vol 259 (9) ◽

pp. 5899-5906 ◽

Cited By ~ 1

Author(s):

S Y Wang ◽

L J Gudas

Keyword(s):

Stem Cell ◽

Retinoic Acid ◽

F9 Teratocarcinoma

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Patterns of Cell Division and the Risk of Cancer

Genetics ◽

10.1093/genetics/163.4.1527 ◽

2003 ◽

Vol 163 (4) ◽

pp. 1527-1532 ◽

Cited By ~ 2

Author(s):

Steven A Frank ◽

Yoh Iwasa ◽

Martin A Nowak

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Basal Layer ◽

Mutation Rates ◽

Tissue Architecture ◽

Cell Lineages ◽

Cell Mutation ◽

Long Stem ◽

Risk Of Cancer ◽

Transit Cell

Abstract Epidermal and intestinal tissues divide throughout life to replace lost surface cells. These renewing tissues have long-lived basal stem cell lineages that divide many times, each division producing one stem cell and one transit cell. The transit cell divides a limited number of times, producing cells that move up from the basal layer and eventually slough off from the surface. If mutation rates are the same in stem and transit divisions, we show that minimal cancer risk is obtained by using the fewest possible stem divisions subject to the constraints imposed by the need to renew the tissue. In this case, stem cells are a necessary risk imposed by the constraints of tissue architecture. Cairns suggested that stem cells may have lower mutation rates than transit cells do. We develop a mathematical model to study the consequences of different stem and transit mutation rates. Our model shows that stem cell mutation rates two or three orders of magnitude less than transit mutation rates may favor relatively more stem divisions and fewer transit divisions, perhaps explaining how renewing tissues allocate cell divisions between long stem and short transit lineages.

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Identification and Characterization of Multiple Myeloma Stem Cell-Like Cells

Cancers ◽

10.3390/cancers13143523 ◽

2021 ◽

Vol 13 (14) ◽

pp. 3523

Author(s):

Wancheng Guo ◽

Haiqin Wang ◽

Peng Chen ◽

Xiaokai Shen ◽

Boxin Zhang ◽

...

Keyword(s):

Multiple Myeloma ◽

Drug Resistance ◽

Stem Cell ◽

New Drugs ◽

Cell Therapies ◽

High Relapse Rate ◽

High Incidence ◽

Poorly Differentiated ◽

Identification And Characterization

Multiple myeloma (MM) is a B-cell tumor of the blood system with high incidence and poor prognosis. With a further understanding of the pathogenesis of MM and the bone marrow microenvironment, a variety of adjuvant cell therapies and new drugs have been developed. However, the drug resistance and high relapse rate of MM have not been fundamentally resolved. Studies have shown that, in patients with MM, there is a type of poorly differentiated progenitor cell (MM stem cell-like cells, MMSCs). Although there is no recognized standard for identification and classification, it is confirmed that they are closely related to the drug resistance and relapse of MM. This article therefore systematically summarizes the latest developments in MMSCs with possible markers of MMSCs, introduces the mechanism of how MMSCs work in MM resistance and recurrence, and discusses the active pathways that related to stemness of MM.

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