- 17-6099-42 - Invitrogen Antibodies CX3CR1 antibody

Submitted references Distinct gene expression patterns for CD14++ and CD16++ monocytes in preeclampsia.

Vishnyakova P, Kuznetsova M, Poltavets A, Fomina M, Kiseleva V, Muminova K, Potapova A, Khodzhaeva Z, Pyregov A, Trofimov D, Elchaninov A, Sukhikh G, Fatkhudinov T
Scientific reports 2022 Sep 14;12(1):15469

Microglia innately develop within cerebral organoids.

Ormel PR, Vieira de Sá R, van Bodegraven EJ, Karst H, Harschnitz O, Sneeboer MAM, Johansen LE, van Dijk RE, Scheefhals N, Berdenis van Berlekom A, Ribes Martínez E, Kling S, MacGillavry HD, van den Berg LH, Kahn RS, Hol EM, de Witte LD, Pasterkamp RJ
Nature communications 2018 Oct 9;9(1):4167

The Transcription Factor Hobit Identifies Human Cytotoxic CD4(+) T Cells.

Oja AE, Vieira Braga FA, Remmerswaal EB, Kragten NA, Hertoghs KM, Zuo J, Moss PA, van Lier RA, van Gisbergen KP, Hombrink P
Frontiers in immunology 2017;8:325

Modulation of Hematopoietic Lineage Specification Impacts TREM2 Expression in Microglia-Like Cells Derived From Human Stem Cells.

Amos PJ, Fung S, Case A, Kifelew J, Osnis L, Smith CL, Green K, Naydenov A, Aloi M, Hubbard JJ, Ramakrishnan A, Garden GA, Jayadev S
ASN neuro 2017 Jul-Aug;9(4):1759091417716610

Dual functions of fractalkine/CX3C ligand 1 in trafficking of perforin+/granzyme B+ cytotoxic effector lymphocytes that are defined by CX3CR1 expression.

Nishimura M, Umehara H, Nakayama T, Yoneda O, Hieshima K, Kakizaki M, Dohmae N, Yoshie O, Imai T
Journal of immunology (Baltimore, Md. : 1950) 2002 Jun 15;168(12):6173-80

Identification and molecular characterization of fractalkine receptor CX3CR1, which mediates both leukocyte migration and adhesion.

Imai T, Hieshima K, Haskell C, Baba M, Nagira M, Nishimura M, Kakizaki M, Takagi S, Nomiyama H, Schall TJ, Yoshie O
Cell 1997 Nov 14;91(4):521-30

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Staining of normal human peripheral blood cells with Anti-Human CD56 (NCAM) PE (Product # 12-0567-42) and Rat IgG2b K Isotype Control APC (Product # 17-4031-82) (left) or Anti-Human CX3CR1 APC (right). Cells in the lymphocyte gate were used for analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2. Microglia-like cells (ScMglia) differentiated from hPSC develop microglia-like morphology and express microglia/myeloid markers. (a, b) Representative images (taken with 4 x objective) of cystic (a) and dense, noncystic (b) EBs in culture at Day 10. (c) Representative phase contrast images at 10 x of differentiating cells at Day 21 and (d) magnified 32 x images of microglial-like precursor cells at Day 21 exhibiting rounded morphologies with microglia-like processes (red box inset in (c)). (e) Phase image (32 x objective) of microglia-like cells (ScMglia) differentiated from iPS cells at Day 28, showing ramified morphology and even spacing among cells that have migrated away from the EB. (f) Most cells with this morphology also stained positively for microglial marker Iba1 (red). (g) Phase imaging showing fewer ScMglia were observed after extended culture (8 to 10 days after P1 plating) and (h) Iba1 immunolabeling demonstrates iPS-derived microglia-like cells continued to be Iba1 immunoreactive. Some microglia-like cells began to exhibit a more rounded morphology. (i) Representative flow cytometry analysis of ScMglia derived from an iPSC line showing 20.02% of gated cells express TREM2 (the M1 population); a total of 4,924 cells (events) were counted. (j) Flow cytometry analysis of ScMglia derived from both iPS and ES cells demonstrates both ES and iPSC-derived microglia-like cells express CX3CR1. (k) Differentiated ScMglia show double immunolabeling for CD11b and C

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunophenotyping of the patient's monocytes. Representative histograms of gated monocytes stained with antibodies to CD40, CD80, CD86, CD163, CD206 or CX3CR1 (magenta-filled histogram) and control samples (empty contour) for three groups of patients ( a ). Percents of positively stained cells are indicated above the gate bars. Levels of CD40+, CD80+, CD86+, CD163+, CD206+ and CX3CR1+ monocytes in three groups: the data are listed as mean +- SD ( b ): eoPE early-onset PE (n = 12), loPE late-onset PE (n = 10), control (n = 11).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 4 oMG expressed microglia-characteristic cell surface markers and showed similar functional immune and phagocytic properties as adult MG. a Flow cytometric analyses of the expression pattern of microglial extracellular markers on CD11b+-gated oMG (oMG 1, 3, and 5) compared to adult MG derived from three separate brain regions from adult MG1.1. (eight organoids were pooled per donor (oMG 1, 3, and 5) after 52 days in culture). b Morphology of magnetic automated cell sorted CD11b+ oMG 1 and adult MG in bright field microscope after 1 week in culture. Scale bar 40 mum. c mRNA expression, determined by qRT-PCR, of pro-inflammatory cytokines IL6 and IL1B after 6 h stimulation with LPS was significantly higher in oMG compared to adult MG (Mann-Whitney test IL6 and IL1B: U = 0, n = 4, p = 0.03). LPS-stimulated response relative to control condition without LPS. ( n = 4 experiments, eight organoids pooled per experiment; adult MG1.1) (* p < 0.05). d Anti-inflammatory response of oMG and adult MG was compared by qRT-PCR for expression of anti-inflammatory genes CD163 and MRC1 upon 72 h stimulation with dexamethasone. Dexamethasone-stimulated response relative to control condition without dexamethasone. (oMG, n = 3 separate experiments in which oMG were isolated from > 4 pooled cerebral organoids from iPSC 1 per experiment; adult MG, n = 4). e Phagocytosis capacity was tested oMG 1 and adult MG by performing a phagocytosis assay with iC3b-coated green-yellow

17-6099-42

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