Macrophages constitute a heterogeneous population of functionally distinct cells involved in several physiological and pathological processes. M1 and m2 macrophages derived from thp1 cells differentially. Monocytes and macrophages are phagocytes, acting in both nonspecific defence or innate immunity as well as specific defense or cellmediated immunity of vertebrate animals. Thp 1 cells show a large, round single cell morphology and express distinct monocytic markers. Differentiated macrophages are the resident tissue phagocytes and sentinel cells of the innate immune response. It is used to test leukemia cell lines in immunocytochemical analysis of proteinprotein interactions, and immunohistochemistry. Mature tissue macrophages convert from circulating monocyte precursor cells by terminal differentiation in a process that is not fully understood. Has anyone got any citation that shows that it is a marker and could. I have several problems activating my thp 1 cells with pma. Given that macrophages play key roles in pathogen recognition, phagocytosis, processing, and antigen presentation, we evaluated the effect of transferon on phenotype and function of macrophagelike cells derived from thp1 monocytes.
Because after facs,my cell surface markers are not coming up. Exosomes from aspc1, an ascitesderived human pdac cell line, were enriched in icam1, which mediated their docking to macrophages through interactions with surfaceexposed cd11c on macrophages. Tg induces upregulation of m1 macrophagespecific markers in. Information about the openaccess article the identification of markers of macrophage differentiation in pmastimulated thp1 cells and monocytederived macrophages. Cell surface activation marker expression on the human openi. Marc daigneault, julie a preston, helen m marriott, moira k b whyte, david h dockrell pmid 20084270. We determined the surface expression of cd80 and cd86 by flow cytometry and il1. The same phenotype was observed in pmadifferentiated thp1 macrophages where ppm1a expression levels increased to 2.
The tumorpromoting effects of pancreatic cancer cell. Jan, 2010 the identification of markers of macrophage differentiation in pmastimulated thp1 cells and monocytederived macrophages. Surface markers are known to predict functional properties. Jan 15, 2020 conservation of myeloid cell markers between humans and bats. Indeed, thp1 cell activations via, for example, mitogens increase cell size as well. Thp1 monocyte to macrophage differentiation and m1 m2. The differentiated thp1 cells, however, yield a macrophageonly. Jan, 2010 surface markers of thp 1 cells during differentiation protocols. Several monocyte29 tomacrophage differentiation protocols exist, with phorbol 12myristateacetate pma. Citeseerx the identification of markers of macrophage. They display remarkable plasticity by changing their phenotype and function in response to environmental cues representing a spectrum of different functional phenotypes.
There was no statistical difference between mdm and pmar thp1 cells. Vd 3 treated cells and monocytes have greater cd14 surface expression in comparison to pmar and mdm. Tg induces upregulation of m1 macrophagespecific markers. Macrophages derived from pma treated thp1 monocytes are very adhesive and they sticked to any surface once activated, any culture t75t25 flaskwell surface are good enough for these macrophages, so theres no need to pruchase extra collagenmatrix treated plate for them. To address this gap in knowledge and as a baseline to compare with tissue and blood monocyte derived macrophages, this study used flow cytometry to examine the surface expression of a wide range of important functional markers on both monocyte and macrophage thp1 cells. Phorbol12myristateacetate pma and 1,25dihydroxyvitamin d3 vd3 are stimuli commonly used to induce macrophage differentiation in monocytic cell lines but the extent of differentiation in comparison to primary tissue macrophages is unclear. Etoposideinduced cancer cell apoptosis was markedly reduced in the presence of thp1 m2 macrophages, while. Aspc1 exosomes also contained much higher levels of arachidonic acid aa, and they fused at a higher rate with thp1derived macrophages than did.
Which are the best differentiation markers in thp1 and u937. Thp1 cells resemble primary monocytes and macrophages in morphology and. For example cd206 is a good m2 marker in mice but cd200r seems to work better in human cells. The pattern of cd marker expression in thp1 and u937 cells was also. B morphological and cell surface marker expression cd11c, cd11b changes observed by microscopic and flow cytometric analysis of pma phorbol 12myrisate acetate differentiated m0. Nearly all thp 1 cells start to adhere to culture plates and differentiate into macrophages after exposure to phorbol12myristateacetate pma, also known as tpa,12otetradecanoylphorbolacetate. Although their markers frequently differ based on their anatomical locations and functions, there is some overlap in the markers that are expressed on different macrophage subpopulations. Well, firstly you should address whether thp1 is becoming a m1 or m2 macrophage. The results indicate that varanadi kashayam reduced the differentiation of thp1 monocytes to macrophages and downregulated the expression of cell surface markers. The analysis of results from both proofofprinciple screening experiments identified 21 markers expressed on both thp1 monocytes and macrophages, 23 markers uniquely expressed on thp1 monocytes, and 20 markers uniquely. Cell surface activation marker expression on the human. Similarities and differences in surface receptor expression. The majority of people that use this cell line are interested in the differentiated macrophage phenotype, which can be achieved by stimulating thp 1 cells.
American type culture collection, manassas, va was used, realizing the cell culture within rpmi 1640 culture media, which was supplemented with 10% inactivated fbs and 1 % antibioticantimycotic life technologies, grand island, ny. Induction of macrophage function in human thp1 cells is. Thp1 cells resemble primary monocytes and macrophages in morphology and differentiation properties. Interleukin32 induces the differentiation of monocytes into. B facs analysis of the expression of the macrophage and dc markers cd14, cd1a, cd64, and cd83 in human pbmcs stimulated for 3 days. On the other hand, several surface markers relevant for differentiating human macrophages from monocytes such as cd163, cd169 and cd206 were observed to have conserved gene homologs in bats. Immunophenotyping monocytes, macrophages and granulocytes. Marc daigneault, julie a preston, helen m marriott, moira k b whyte, david h dockrell. I tried to look for cd14 cell surface marker on thp1 cells and could not see them. Ebscohost serves thousands of libraries with premium essays, articles and other content including the identification of markers of macrophage differentiation in pmastimulated thp1 cells and monocytederived macrophages. Cd80 expression correlates with il6 production in thp1like. I use pma to differentiate thp1 and u937 cells to macrophage.
Find your cell specific markers using our mouse or human immune cell lineage and expression guide, posters and interactive database. Interleukin32 induces the differentiation of monocytes. Arginine transport in human monocytic leukemia thp. Immunophenotyping monocytes, macrophages and granulocytes in.
Terminal differentiation can be monitored by changes in morphology, adherence, phagocytosis, expression of surface markers, or the release of pge and tnf 24, 25. Macrophages are key in orchestrating immune responses to microenvironmental stimuli, sensed by a complex set of surface receptors. In summary, pmadifferentiated thp1 macrophages did not entirely. The mannose receptor, cd206, a prototypic mouse m2 marker, is highly expressed in mcsftreated macrophages in mouse, whereas in human monocytes cd206 is substantially induced by gmcsf 9, 10. Thp1 cells and identify increased numbers of mitochondria and lysosomes, resistance to apoptosis and the potency of tlr2. Jan 17, 2018 the same phenotype was observed in pmadifferentiated thp1 macrophages where ppm1a.
For several reasons, monocytic cell lines such as thp1 have been considered as a. Thp1 cells show a large, round singlecell morphology and express distinct monocytic markers. In terms of phenotypic surface markers, while m1 polarization led to the expected outcome in both cells types, m2 polarization did not seem to affect thp1 macrophages. Recently, we found that triglyceride tgtreated thp1 monocytes express a variety of macrophagespecific surface markers, indicating that tg treatment could trigger the differentiation of monocytes into macrophages. The human cell line thp1 has a monocytic phenotype, including the ability to differentiate into macrophages, providing a tractable, standardised surrogate for human monocytederived macrophages. I usually use cd11b as differentiation marker but i need another one. Plos one 2010120 the identification of markers of macrophage differentiation in pmastimulated thp1 cells and monocytederived macrophages. As a model for monocytes, the thp 1 cell line number tib202l. To identify potential surface markers for experimental validation, we first evaluated bat genomes two each from yinpterochiroptera. Tlr2 is another surface marker that is downregulated with macrophage differentiation. The identification of markers of macrophage differentiation in pmastimulated thp1 cells and monocytederived macrophages. The identification of markers of macrophage differentiation.
Metabolic characterisation of thp1 macrophage polarisation. Which are the best differentiation markers in thp1 and u937 cells. The phenotype of mature tissue macrophages represents the composite of environmental and differentiationdependent imprinting. Compared with primary cells, the response of thp 1 to lps stimulation and to oxidized lowdensity lipoproteins was attenuated, and tolllike receptor tlr 3 and tlr5 mrna expression was lower. In conclusion, tg enhances the expression of m1 macrophage markers including cd40, cd80 and cd86 in thp1 cells and decreases the expression of m2 macrophage markers, cd200r. Cell surface marker expression with macrophage differentiation. Similarities and differences in surface receptor expression by thp1. May 17, 2011 terminal differentiation can be monitored by changes in morphology, adherence, phagocytosis, expression of surface markers, or the release of pge and tnf 24, 25.
Cell culture and differentiation thp1 cells, derived from a human monocytic leukaemia, were grown in rpmi 1640 supplemented with 15% foetal calf serum fcs. Which are the best differentiation markers in thp1 and. Thp1 differentiated macrophages expressed a wider repertoire of receptors. The phosphatase ppm1a controls monocytetomacrophage. This study aimed to characterise m1 and m2 phenotypes in thp1 macrophages in order to identify characteristic metabolites of each polarisation state. Cytotoxicity assessment, inflammatory properties, and. Furthermore, a subset of pmatreated thp 1 cells does not express scavenger receptors, demonstrating heterogeneity in this cell line. Dec 31, 2016 in conclusion, tg enhances the expression of m1 macrophage markers including cd40, cd80 and cd86 in thp1 cells and decreases the expression of m2 macrophage markers, cd200r. Is there any specific markers that can be used to confirm the differentiation process, or that.
Frontiers induction of macrophage function in human thp1. The expression of the surface markers that define mouse m1 and m2 macrophages is different in humans. Screen suspension thp1 cells monocytes stained with the bd lyoplate human cell surface marker screening panel using flow cytometry. So, you should stain with cd206 for m2 subtype and cd1632 m1 subtype. This argues against a role of plasma in induction of thp1 cell differentiation. The thp1 cell line is a human monocytic leukaemia cell line which was established in 1980 by tsuchiya et al. I expected cd 14 on the cells as they are supposedly monocytes. Cell surface activation marker expression on the human thp1 pma differentiated macrophages for mhci and mhcii molecules a and costimulatory cd80, cd86 and cd40 molecules b is modulated by enprocal d and compared with other digested milk product controls in vitro. The socalled m1 and m2 macrophages are often considered as representative. Based on these results, we suggest that tg affects differentiation of monocytes into m1 macrophages which may be involved in proinflammatory responses in the development. And ppl say that we shouldnt use trypsin,so what else can we use to lift the cells up from the surface.
Dockrell, title the identification of markers of macrophage differentiation in pmastimulated thp1 cells and, year. Etoposideinduced cancer cell apoptosis was markedly reduced in the presence of thp1 m2 macrophages, while apoptosis was increased in cells cocultured. Standardized protocols for differentiation of thp1 cells. Frontiers convenience versus biological significance. Cd14 is a monocyte marker that is downregulated during differentiation. Thp1 cell surface markers for flow cytometry flow cytometry. Macrophages are found throughout the body in all tissues where they have. Comparative genotypic and phenotypic analysis of human. Jun 11, 2019 in this article, orlova and colleagues show that monocytes and macrophages differentiated from human induced pluripotent stem cells hipscs are functionally comparable with peripheral blood pbderived cells, although some differences were identified. Monocytes and macrophages are phagocytes, acting in both nonspecific defence or innate immunity as well as specific defense or cellmediated immunity of. Furthermore, it could decrease the release of proinflammatory cytokines from lpsinduced thp1 macrophages and downregulated the expression of tnf.
A schematic representation of treatments to differentiate thp1 monocytes mo into macrophages m0 or polarized macrophages m1, m2. Thp1 cell surface markers for flow cytometry need a robust marker for undifferentiated thp1 cells sep242009 i am working with thp1 cells atcc tib 202, a human monocytic cell line. Flow cytometry and highcontent imaging to identify. In order to obtain activated monocytes, thp 1 cells. Are there any specific cell surface markers for m1 and m2. Phorbol12myristateacetate pma and 1,25dihydroxyvitamin d3 vd3 are.
Nearly all thp1 cells start to adhere to culture plates and differentiate into macrophages after exposure to phorbol12myristateacetate pma, also known as tpa,12otetradecanoylphorbolacetate. Thp 1 cell surface markers for flow cytometry need a robust marker for undifferentiated thp 1 cells sep242009 i am working with thp 1 cells atcc tib 202, a human monocytic cell line. American journal of respiratory cell and molecular biology. Citeseerx document details isaac councill, lee giles, pradeep teregowda. Macrophages represent the primary human host response to pathogen infection and link the immediate defense to the adaptive immune system. Cd14 will stain either macrophage subtypes and is expressed even in undifferentiated human monocytes upon activation you may see cd14 upregulation. The majority of people that use this cell line are interested in the differentiated macrophage phenotype, which can be achieved by stimulating thp1 cells. In the present study, thp1 macrophages were found to be more sensitive to nx nps than heladerived epithelial cells as no ctb reduction was observed in tzmbl cells treated with 35 gml of nx nps for 48 h. However, it is unclear how the differentiation state of pmatreated thp1 cells compares with that of tissue macrophages or monocytederived macrophages. Objective to investigate the effect of chinese prepared medicine naoxintong on scavenger receptors cd36 and sra expression of human thp1 monocytederived macrophages.
Flow cytometry and highcontent imaging to identify markers. The markers were also evaluated on the cell surface of the mdms obtained from 11 to 16 ssc patients not treated with glucocorticoids and compared with those obtained from hss. M1 macrophages promote inflammatory responses, whereas m2 macrophages exhibit antiinflammatory activity. Differentiated adherent thp1 macrophages were screened using a bd pathway highcontent imaging system. All the facts at your fingertips from cell lineage to marker proteins, discover for yourself. Published protocols using the promonocytic cell line thp 1 have tended to result in cells that closely resemble classicallyactivated macrophages, differentiated in ifn. A rtpcr of the macrophage and dc cell markers cd1a, cd14, and cd64 after stimulation of pbmcs with 30 ngml il32. In this article, orlova and colleagues show that monocytes and macrophages differentiated from human induced pluripotent stem cells hipscs are functionally comparable with peripheral blood pbderived cells, although some differences were identified.
Here, we analyzed the protein kinases of the human monocytic cell line thp1 before and after induction of. Cd80 expression correlates with il6 production in thp1. Tumorpromoting effects of pancreatic cancer cell exosomes. Dclike morphology and surface marker expression in u937 stephens et al. Pma withdrawal in pmatreated monocytic thp1 cells and. Vd 3 treated cells and monocytes have greater cd14 surface expression in comparison to pmar and mdm figure 3a. Analysis of cd206 plasma membrane expression by flow cytometry.
The identification of markers of macrophage differentiation in pma. Furthermore, it is well known that the cytotoxicity potential of a given compound varies with the cell type. I tried to look for cd14 cell surface marker on thp1. We have compared the phenotype of the promonocytic thp1 cell line after various protocols of differentiation utilising vd3 and pma in comparison to primary human monocytes or monocytederived macrophages mdm. Common macrophage markers tissueresident macrophages are a heterogeneous group of cells that express a variety of cell surface and intracellular markers. Herein, untargeted liquid chromatography lcmass spectrometry msbased metabolite profiling was applied to characterise the metabolic profile of m1like and m2like thp1 macrophages. Responses were measured by the upregulation of the surface markers cd80, cd86 and cd40. Thp1 cell line culture and stimulation as a model for monocytes, the thp1 cell line number.
I tried to look for cd14 cell surface marker on thp 1 cells and could not see them. The phorbol 12myristateacetate differentiation protocol is critical to the interaction of thp1 macrophages with salmonella typhimurium the identification of markers of macrophage differentiation in pmastimulated thp1 cells and monocytederived macrophages. Yet, mitogenactivated and thp1 cellderived macrophages display a distinct surface marker signature e. Surface markers of thp1 cells during differentiation protocols. Redox stimulation of human thp1 monocytes in response to. Phorbol12myristateacetate pma and 1,25dihydroxyvitamin d3 vd3 are stimuli commonly used to induce macrophage differentiation in monocytic cell. It was derived from the blood of a patient with acute monocytic leukaemia. For example both m1 and m2 will express mhcii, but m1 will express it with much greater intensity. In vitro models of differing macrophage functions are useful since human monocytederived macrophages are shortlived, finite and vary from donor to donor. Daigneault m1, preston ja, marriott hm, whyte mk, dockrell dh. Specifically, not only cd206 expression did not differ between m1 and m2 cells, but there was a trend toward increased cd206 levels in m1 in comparison to m2 thp1 macrophages. Thp 1 is a human monocytic cell line derived from an acute monocytic leukemia patient.
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