Journal: Biosensors

Article Title: White Light Spectroscopy Characteristics and Expansion Dynamic Behavior of Primary T-Cells: A Possibility of Online, Real-Time, and Sampling-Less CAR T-Cell Production Monitoring

doi: 10.3390/bios15040251

Figure Lengend Snippet: List of reagents and equipment.

Article Snippet: 7-AAD Viability Staining Solution , SONY Biotechnology (San Jose, CA, USA, supplier France) , 2702020.

Techniques: Staining, Cytometry, Flow Cytometry, Software

Journal: Nature

Article Title: Neonatal imprinting of alveolar macrophages via neutrophil-derived 12-HETE

doi: 10.1038/s41586-022-05660-7

Figure Lengend Snippet: (a) Representative FACS plot for the determination of AM progenitors (pre-AM and fetal liver monocytes) and AM in the lungs of PND0, PND1 and PND3 WT and Alox15 −/− mice. (b) Weight of lungs from PND1 and PND3 WT (n = 7/group) and Alox15 −/− mice (n = 6/group). (c-f) Lungs from PND1 or PND3 WT and Alox15 −/− mice were harvested and the levels of TGF-β1 (c) (PND1, WT: n = 6, Alox15 −/− : n = 7; PND3, WT: n = 8, Alox15 −/− : n = 7), GM-CSF (d) (PND1, WT: n = 6, Alox15 −/− : n = 7; PND3, WT: n = 8, Alox15 −/− : n = 8), type 1 and type 2 cytokines (e- PND1: WT: n = 6, Alox15 −/− : n = 7, f- PND3: n = 8/group) were determined by ELISA. (g-h) BAL-AM were isolated from PND3 WT and Alox15 −/− mice for gene expression analysis (n = 3 biological replicates/group). (i) BAL-AM were isolated from adult naïve animals and cell death was assessed by flow cytometry using AnnexinV/7AAD staining (n = 7/group). (j-l) Batch effect examinations. (j) PCA and (k) heatmap plots that suggest WT4 is an outlier. (l) A heatmap that indicates the similarity between different samples. Data are presented as mean ± s.e.m and are pooled from two (b-f, i) or three (g-h) independent experiments.

Article Snippet: Necrosis and apoptosis levels of BAL AMs were assessed using a PE-AnnexinV and 7-amino-actinomycin D (7-AAD) Apoptosis Detection Kit I (BD Biosciences) according to the manufacturer’s instructions and analysed by flow cytometry.

Techniques: Enzyme-linked Immunosorbent Assay, Isolation, Expressing, Flow Cytometry, Staining

Journal: bioRxiv

Article Title: Human skeletal muscle CD90 + fibro-adipogenic progenitors are associated with muscle degeneration in type 2 diabetic patients

doi: 10.1101/2020.08.25.243907

Figure Lengend Snippet: A-i , Contour flow-plots from the sorting strategy identifying cell negative for CD45, CD31 and Propidium Iodide (PI) and either CD34 + CD56 − or CD34 − ; A-ii , displaying a CD34 + CD90 − and a CD34 + CD90 + population; A-iii , displaying a CD56 + CD82 + and a CD56 − CD82 − population; A-iv , displaying a CD90 + and a CD90 − population; B , FACS work flow for cell characterization; C-E , adipogenic, fibrogenic and myogenic differentiation of CD34 + CD90 − CD56 − ( C ), CD34 + CD90 + CD56 − ( D ) and CD56 + CD82 + CD34 − ( E ) populations verified by immunocytochemistry for Perilipin-1 (Red) and Collagen-1 (green) or Desmin (Red) and Myosin Heavy Chain (MyHC, green); F , Content of mononuclear cells in human skeletal muscle (n=17) in percent of the total number of single cells; G , Expression of PDGFRa in freshly sorted FAPs; H , Distribution of COL1A1 mRNA expressing CD90 + CD56 − (Fibro-adipogenic progenitors; FAPs), CD56 + (Muscle stem cells; MuSCs) and CD90 − CD56 − (Un-identified) cells in human skeletal muscle (n=6) after activation for 3-9 days in vitro; I , Work-flow for single-cell clonal experiment; J , Colony formation (percentage wells containing colonies) of single sorted FAPs and MuSCs (n=12); K , Limiting dilution assay of sorted FAPs and MuSCs (n=3) with fraction of negative wells as a function of deposited cells. Solid line represents non-linear fit; L , MyHC and Desmin positive myotubes from clonal cultures from single cell sorted MuSCs (stitched images from entire well); M , Adipogenic (Perilipin-1 + ) and fibrogenic (Collagen-1 + ) cells in clonal cultures from single cell sorted FAPs (stitched images from entire well); N , GO: Biological processes enriched in freshly sorted FAPs; O , Percentage of MuSCs and FAPs incorporating EdU (S-phase entry) post isolation (n=10-16); P , Images of EdU incorporation 96h post sort of MuSCs ( i ) and FAPs ( ii ). Significant difference denoted by *p<0.05, **p<0.01 and ***p<0.001.

Article Snippet: 130-101-302 10μl/sample, Miltenyi Biotec) and CD34-APC (20μl/sample, clone 581, BD Bioscience) in darkness at 4°C for 30 min. Propidium iodide (PI, 10μl/sample, cat 556463, BD Bioscience) immediately before sorting to exclude non-viable cells.

Techniques: Immunocytochemistry, Expressing, Activation Assay, In Vitro, Limiting Dilution Assay, Isolation