Phenotype and functional identity of GM-CSF–independent dendritic cells generated by long-term propagation of DC progenitor cells in bone marrow cells and skin langerhans cells

Katayama, Hokahiro; Hattori, Yuuichi; Ogata, Keiichi; Yan, Hua; Satoh, E.; Teramoto, Koji; Arii, Shigeki; Kamide, Ryoichi; Nakagawa, Hidemi; Kimura, Hiroyuki

doi:10.1016/j.transproceed.2005.02.017

Cited by 4 publications

(3 citation statements)

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“…Several developmental progenitors and precursors of conventional DC have been identified, including MDP (monocyte-macrophage dendritic progenitor) [ 19 – 21 ] and CDP (common dendritic precursor) [ 22 – 25 ]. In the developmental pathway of GM-CSF-driven or monocyte-derived DC (moDC), the early stages of development include common myeloid progenitors (CMP) [ 26 , 27 ], which give rise to granulocyte macrophage progenitors (GMP) [ 15 , 28 ], followed by monocytes. A precursor of monocytes and macrophages but not dendritic cells (cMOP) has also been identified in the bone marrow [ 19 ], yet its place in the GM-CSF-driven differentiation pathway remains to be determined.…”

Section: Introductionmentioning

confidence: 99%

Analysis of the developmental stages, kinetics, and phenotypes exhibited by myeloid cells driven by GM-CSF in vitro

2017

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The developmental progression of conventional DC has been quite well defined, yet the developmental pathway of monocyte-derived, GM-CSF-driven DC is less well understood. We addressed this issue by establishing an isolation strategy that identifies five distinct GM-CSF derived cell types. Expression of Ly6C and CD115 (Csf-1R) was used to identify and isolate four populations. One of the populations could be further separated based on CD11c expression, distinguishing five populations. We further defined these cells based on expression of transcription factors and markers of early and later stages of myeloid development. These discreet developmental stages corresponded well with previously defined populations: Common Myeloid Progenitors (CMP), Granulocyte/Macrophage Progenitors (GMP), Monocytes, as well as Monocyte-derived macrophages (moMac) and Monocyte-derived DC (moDC). Finally, within the moMac population we also identified moDC precursor activity (moDP) that could be distinguished from moMac and moDC based on their level of MHC class II expression and developmental plasticity.

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Section: Introductionmentioning

confidence: 99%

Analysis of the developmental stages, kinetics, and phenotypes exhibited by myeloid cells driven by GM-CSF in vitro

2017

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“…There is an increasing appreciation that understanding biological decision‐making requires tracing information flow through cells, which necessitates assaying multiple measures of genotype and phenotype on hundreds or thousands of individual cells. Additionally, there is inherent functional heterogeneity among cell types, among single cells within defined cell types, and even among clonal populations; this heterogeneity holds significant promise for elucidating the mechanisms of many processes in health and disease and thus motivates the development of assays applicable to sparse populations of cells (defined as single cells or groups of 2–10 cells) . Novel platforms that have been developed to achieve this goal often center on microfluidic approaches, such as arrays of micro/nano/picoliter‐volume wells, microchambers, and emulsion‐based droplet technologies .…”

Section: Introductionmentioning

confidence: 99%

Stochastic Particle Barcoding for Single-Cell Tracking and Multiparametric Analysis

Castellarnau

Szeto

et al. 2014

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This study presents stochastic particle barcoding (SPB), a method for tracking cell identity across bioanalytical platforms. In this approach, single cells or small collections of cells are co-encapsulated within an enzymatically-degradable hydrogel block along with a random collection of fluorescent beads, whose number, color, and position encode the identity of the cell, enabling samples to be transferred in bulk between single-cell assay platforms without losing the identity of individual cells. The application of SPB was demonstrated for transferring cells from a subnanoliter protein secretion/phenotyping array platform into a microtiter plate, with re-identification accuracies in the plate assay of 96±2%. Encapsulated cells were recovered by digesting the hydrogel, allowing subsequent genotyping and phenotyping of cell lysates. Finally, a model scaling was developed to illustrate how different parameters affect the accuracy of SPB and to motivate scaling of the method to 1,000's of unique blocks.

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“…The cellular phenotype is a conglomeration of multiple cellular processes, representing varying expression levels of genes and proteins that determine the cell's particular function in activities such as cellular communication, adhesion, or metabolism 1 2 3 4 . Average measurements of such molecular markers in cell populations obscure inherent cell-cell heterogeneity and restrict the ability to distinguish between the responses of individual cells within a sample 2 5 6 7 . While flow cytometry has traditionally been used to determine single cell phenotypes, it cannot provide continuous measurements of proteins in the same individual cells over time 7 .…”

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confidence: 99%

Live single cell functional phenotyping in droplet nano-liter reactors

Konry

Golberg

Yarmush

2013

Sci Rep

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While single cell heterogeneity is present in all biological systems, most studies cannot address it due to technical limitations. Here we describe a nano-liter droplet microfluidic-based approach for stimulation and monitoring of surfaceand secreted markers of live single immune dendritic cells (DCs) as well as monitoring the live T cell/DC interaction. This nano-liter in vivo simulating microenvironment allows delivering various stimuli reagents to each cell and appropriate gas exchanges which are necessary to ensure functionality and viability of encapsulated cells. Labeling bioassay and microsphere sensors were integrated into nano-liter reaction volume of the droplet to monitor live single cell surface markers and secretion analysis in the time-dependent fashion. Thus live cell stimulation, secretion and surface monitoring can be obtained simultaneously in distinct microenvironment, which previously was possible using complicated and multi-step in vitro and in vivo live-cell microscopy, together with immunological studies of the outcome secretion of cellular function.

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Phenotype and functional identity of GM-CSF–independent dendritic cells generated by long-term propagation of DC progenitor cells in bone marrow cells and skin langerhans cells

Cited by 4 publications

References 4 publications

Analysis of the developmental stages, kinetics, and phenotypes exhibited by myeloid cells driven by GM-CSF in vitro

Analysis of the developmental stages, kinetics, and phenotypes exhibited by myeloid cells driven by GM-CSF in vitro

Stochastic Particle Barcoding for Single-Cell Tracking and Multiparametric Analysis

Live single cell functional phenotyping in droplet nano-liter reactors

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