Expression Of Marker Molecules Research Articles

Effects of Platelet-Rich Plasma on Macrophage Phenotype and Its Influencing Factors

To evaluate the effects of platelet-rich plasma (PRP) supernatants with different activation methods and storage time on human monocyte-derived macrophages phenotype and explore the possible mechanism. Human monocyte-derived macrophages were cultured in vitro with PRP or activated PRP supernatants activated with different activators. The expression of marker molecules on the surface of macrophages was detected by flow cytometry, and the concentration of growth factors in PRP supernatants was detected by ELISA. After 24 h of coculture, the expression level of CD86 in macrophages stimulated by PRP supernatant (thrombin and Cacl2 activated) was significantly higher than that by PRP group (P<0.05), and the expression of CD163 in macrophages was increased by Cacl2 activated PRP supernatant. Compared with different activator groups, the expression of CD163 in macrophages of Cacl2 activated group was significantly higher than that of thrombin and ADP groups (P<0.05). ELISA results showed that the concentrations of FGF (P<0.001) and EGF (P<0.05) in the supernatant of PRP stored at -80 ℃ for more than 20 months and 10-20 months were significantly higher than those in the group stored at less than 10 months after Cacl2 activation, and the expressions of CD86 (P<0.01), CD163 (P<0.001) and CD206 (P<0.001) in macrophages cocultured with the supernatant of the two groups were significantly increased. PRP activated by different activators has different effects on the phenotype of macrophages. Meanwhile, the storage time will also affect the growth factor concentration and effect of PRP.

Time course of structural and functional maturation of human olfactory epithelial cells in vitro

The unique ability of olfactory neurons to regenerate in vitro has allowed their use for the study of olfactory function, regeneration, and neurodegenerative disorders; thus, characterization of their properties is important. This present study attempts to establish the timeline of structural (protein expression) and functional (odorant sensitivity) maturation of human olfactory epithelial cells (hOE) in vitro using biopsy-derived cultured tissue. Cells were grown for 7 days; on each day, cells were tested for odorant sensitivity using calcium imaging techniques and then protein expression of each cell was tested using immunocytochemistry for proteins typically used for characterizing olfactory cells. Previous studies have shown that mature olfactory neurons in vitro attain a unique "phase-bright" morphology and express the olfactory marker protein (OMP). By day 3 in vitro, a variety of cells were odorant-sensitive, including both "phase-bright" and "phase-dark" cells that have previously been considered glial-like cells. The functional maturation of these hOEs appears to take place within 4 days. Interestingly, the emergence of an odorant sensitivity profile of both phase-bright and phase-dark cells preceded the expression of marker protein expression for OMP (which is expressed only by mature neurons in vivo). This structural maturation took 5 days, suggesting that the development of odorant sensitivity is not coincident with the expression of marker molecules that are hallmarks of structural maturation. These results have important implications for the use of hOEs as in vitro models of olfactory and neuronal function.

Unveiling cell phenotypes in lung vascular remodeling

PULMONARY VASCULAR REMODELING is a hallmark of most forms of pulmonary hypertension (PH), both primary and secondary. This remodeling takes the form of concentric medial thickening of small arterioles, neomuscularization of previously nonmuscular capillary-like vessels, and structural wall changes in larger pulmonary arteries. The pulmonary arterial muscularization is characterized by increased numbers of vascular smooth muscle cells (SMCs) as well as by hypertrophy of individual SMCs (25). In human PH, characteristic plexiform lesions develop, and there is increasing discussion of neovasculogenesis and angiogenesis during remodeling. The vascular remodeling is associated with hypoxia- or inflammationinduced production of growth factors, angiogenic factors, inflammatory mediators, and vasoconstrictors. For decades, investigators have attempted to dissect the mechanisms behind this lung-specific vascular remodeling, investigating a wide range of environmental stimuli, hemodynamic events, biochemical and molecular signaling pathways, and extracellular matrix changes. Recently, investigators have begun looking in greater detail at the heterogeneity of cells in lung vascular walls under normal and pathological remodeling conditions (14, 26). Initially, it was recognized that even within a single region, endothelial cells differ and may have highly distinct characteristics and functions. Now there is increasing interest in and evidence for the existence of diverse cell types within the media of remodeling lung vessels. Emerging evidence suggests that endogenous or circulating inflammatory and/or progenitor cells contribute significantly to the remodeling process (9, 22). The term “circulating inflammatory/progenitor cells” characterizes a wide variety of cell populations that differ with respect to their structural characteristics, expression of marker molecules, and biological functions. These cells include macrophages and other inflammatory cells, mononuclear cells, stem cells, endothelial progenitor cells, fibrocytes, and myofibroblasts. The nature of these cells, their relative importance, and their derivation and source continue to be unclear and highly controversial. The phenotype of these cells may change during the remodeling process or vary with the specific initiating pathophysiology or type of PH. Thus, it is important to define the phenotype of cells in the vascular media and the roles of the cells in the normal and diseased pulmonary vasculature. The exact source of the cells is also important to know because it may determine cell phenotype and targets for

Novel conserved oligodendrocyte surface epitope identified by monoclonal antibody 4860

Oligodendrocytes are the myelinating cells of the central nervous system. They differentiate from oligodendrocyte precursor cells through several intermediate states that can be followed by characteristic morphological changes and the expression of marker molecules. However, most oligodendrocyte lineage markers demarcate either the precursor or the differentiated oligodendrocyte in restricted subcellular compartments. Here, we describe a novel marker of the oligodendrocyte lineage recognised by the monoclonal antibody clone 4860. It selectively labels the surfaces of differentiated oligodendrocytes in culture and clearly differs from other oligodendrocyte markers. Importantly, the 4860 epitope highlights developing white matter tracts in rodent and avian brains and thus represents a useful and conserved feature. The 4860 epitope is not associated with protein backbones as revealed by the related 487/L5 antibody. Furthermore, the epitope disappears upon lipid extraction from cryosections or inhibition of sphingolipid synthesis in cultured oligodendrocytes. Thus, we conclude that mAb 4860 represents a novel lipid-based oligodendrocyte marker.

Effect of Dexamethasone on the Surface Expression of Marker Molecules and Differentiation of Murine B Cells

Background: There are at least two different subsets of B cells, B-1 and B-2. The characteristic features and function of B-2 cells in addition to the effect of steroids on B-2 cells are well-known. Although B-1 cells have different features and functions from B-2 cells, the effect of steroids on B-1 cells is not completely understood. Therefore, this study examined the effects of dexamethasone on peritoneal (or B-1 cells) and splenic B cells (or B-2 cells). Methods: Purified B cells were obtained from the peritoneal fluid and the spleens of mice. The isolated B cells were cultured in a medium and after adding different concentrations of dexamaethasone. The cell survival rate was measured by flow cytometry using propidium iodide. The expression level of the B cell surface marker was analyzed by flow cytometry. During the culture of these cells, immunoglobulin secreted into the culture supernatants was evaluated by an enzyme-linked immunosorbent assay. Results: The survival rate of peritoneal and splenic B cells decreased with increasing dexamethasone concentration. However, the rate of peritofieal B cell apoptosis was lower than that of splenic B cells. CDS and B7.1 expression in peritoneal B cells and CD23 and sIgM expression in splenic B cells after the dexamethasone treatment were reduced. When B cells were treated with dexamethasone, the spontaneous IgM secretion decreased with increasing dexamethasone concentration. Conclusion: Dexamethasone induces apoptosis in peritoneal and splenic B cells. However, peritoneal B cells are less sensitive to dexamethasone. The dexamethasone suppressed expression of the surface markers in peritoneal B cells is different from those in splenic B cells.

The First Prospective Human Evidence That Low Numbers of Circulating Endothelial Precursor Cells Predict Future Cardiovascular Events-Evidence of Defective Vascular Repair?: Reduced Number of Circulating Endothelial Progenitor Cells Predicts Future Cardiovascular Events: Proof of Concept for the Clinical Importance of Endogenous Vascular Repair. Circulation 111: 2981-2987, 2005.

The First Prospective Human Evidence That Low Numbers of Circulating Endothelial Precursor Cells Predict Future Cardiovascular Events-Evidence of Defective Vascular Repair?: Reduced Number of Circulating Endothelial Progenitor Cells Predicts Future Cardiovascular Events: Proof of Concept for the Clinical Importance of Endogenous Vascular Repair. Circulation 111: 2981-2987, 2005.

Simultaneous Detection of Multiple Bone-Related mRNAs and Protein Expression during Osteoblast Differentiation: Polymerase Chain Reaction and Immunocytochemical Studies at the Single Cell Level

Messenger RNA expression analyzed by in situ hybridization and Northern analysis and protein expression analyzed biochemically or immunocytochemically have been used to study the developmental expression of various osteoblast (OB)-associated molecules. These approaches have shown that over a time course of OB differentiation in vivo and in vitro, the expression of macromolecules associated with OB cells changes. However, ambiguities in data from different approaches and in populations representative of cells at different developmental stages are extant. To begin to discriminate differentiation stages with more precision and to address intercellular heterogeneity, fetal rat calvaria cells were grown at low densities under conditions in which bone nodules form and mineralize and colonies were classified morphologically as fibroblastic or osteoblastic (early, intermediate, or mature). Whole discrete colonies and single cells from individual colonies were analyzed molecularly by a random amplification poly(A)-polymerase chain reaction (PCR) and for protein expression by immunocytochemistry; we analyzed the expression of known bone-related macromolecules (collagen type I, alkaline phosphatase, osteopontin, bone sialoprotein, and osteocalcin). Both PCR and immunocytochemistry revealed that different colony types were reproducibly distinguishable in their expression of either general (collagen type I) or bone-associated (alkaline phosphatase, osteopontin, bone sialoprotein, and osteocalcin) macromolecules, such that fibroblastic colonies were distinguishable from osteoblastic colonies and the latter could be subdivided into less mature or more mature osteoblastic colonies. While some aspects of the temporal differentiation sequence defined earlier were confirmed, several additional features were evident from these single cell-single colony studies. First, different repertoires of OB-associated markers were expressed in different cells, suggesting variation in the switch-on of the OB differentiation program and heterogeneity in the OB phenotype. Second, among colonies classified as fibroblastic on the basis of morphology heterogeneity was also evident and there were some cells expressing features consistent with their being osteoprogenitor cells. Our data support the hypothesis that individual fibroblastic and osteoblastic cells are heterogeneous in expression of marker molecules. We also conclude that individual cells and colonies analyzed by poly(A)-PCR will be useful in lieu of mass populations to extend investigation of stages in the progression of OB differentiation.