Journal: Applied Materials Today

Article Title: 3D bioprinting of multilayered scaffolds with spatially differentiated ADMSCs for rotator cuff tendon-to-bone interface regeneration

doi: 10.1016/j.apmt.2022.101510

Figure Lengend Snippet: Fig. 3. Identification and differentiation of rabbit ADMSCs. (A) Harvested adipose tissue from rabbit inguinal region (~ 2 g). (B) Isolated rabbit ADMSCs (scale bar: 200 µm). (C) The surface marker expression percentages of CD34, CD44, CD45, CD90 clone OX-7, and CD105 were revealed by flow cytometry. gray histograms depict isotype labeling for the associated antibodies. (D & E) The gene expressions of TNMD and MKX for verifying the tenogenic differentiation of rabbit ADMSCs. (F & G) The gene expressions of COL2A1 and ACAN for verifying the chondrogenic differentiation of rabbit ADMSCs. (H & I) The gene expressions of RUNX2 and BMP2 for verifying the osteogenic differentiation of rabbit ADMSCs. (n = 6, *p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: We used the following primary antibodies: CD34 (MEC14.7, phycoerythrin (PE), BioLegend), CD44 (W4/86, purified, Bio-Rad), CD45 (L12/201, purified, Bio-Rad), CD51 (23C6, purified, BioLegend), CD90 (OX-7, PE, BioLegend), CD90 (5E10, purified, BioLegend), CD105 (SN6, PE, Invitrogen), rat IgG2a isotype (RTK2758, PE, BioLegend), mouse IgG1 isotype (MOPC-21, purified, BioLegend), and mouse IgG1 isotype (MOPC-21, PE, BioLegend).

Techniques: Isolation, Marker, Expressing, Flow Cytometry, Labeling

Journal: Magnetochemistry

Article Title: Enrichment of Rabbit Primitive Hematopoietic Cells via MACS Depletion of CD45+ Bone Marrow Cells

doi: 10.3390/magnetochemistry7010011

Figure Lengend Snippet: Figure 1. Flow cytometric strategy for the evaluation of magnetic-activated cell sorting (MACS) sorting efficiency. Firstly, live bone marrow mononuclear cells (BMMCs) were gated and then the number of single (CD45+) and double (CD45+LCR+) positive cells in control unsorted samples (A), negative (B), or positive (C) fractions was measured. Gating on CD45+ cells in control and both fractions (D, E, and F, respectively) shows the location of CD45−cells on FSC/SSC dot-plot as well as clear enrichment of these cells in negative fraction compared to control sample or positive fraction. BMMCs—bone marrow mononuclear cells, LCR—labelling check reagent.

Article Snippet: Cells were firstly stained with anti-rabbit CD45 monoclonal antibody (L12/201, mouse IgG1; Bio-Rad, Hercules, CA, USA), then co-labeled with anti-mouse IgG1 MicroBeads (Miltenyi Biotec, Bergisch Gladbach, Germany) and sorted by AutoMACS Pro Separator (Miltenyi Biotec, Bergisch Gladbach, Germany) using two sorting strategies (Posselds and Depl025) as described previously [22].

Techniques: FACS, Control

Journal: Magnetochemistry

Article Title: Enrichment of Rabbit Primitive Hematopoietic Cells via MACS Depletion of CD45+ Bone Marrow Cells

doi: 10.3390/magnetochemistry7010011

Figure Lengend Snippet: Figure 4. Evaluation strategy for the phenotyping of cultured CD45−cells. Firstly, live cultured cells were gated, then only CD45−cells were shown on FSC/SSC dot-plot (A) and cells from this region were analyzed for the expression of specific markers. (B) Blue: control, red: specific marker.

Article Snippet: Cells were firstly stained with anti-rabbit CD45 monoclonal antibody (L12/201, mouse IgG1; Bio-Rad, Hercules, CA, USA), then co-labeled with anti-mouse IgG1 MicroBeads (Miltenyi Biotec, Bergisch Gladbach, Germany) and sorted by AutoMACS Pro Separator (Miltenyi Biotec, Bergisch Gladbach, Germany) using two sorting strategies (Posselds and Depl025) as described previously [22].

Techniques: Cell Culture, Expressing, Control, Marker

Journal: Nature Communications

Article Title: Intestinal intraepithelial lymphocyte activation promotes innate antiviral resistance

doi: 10.1038/ncomms8090

Figure Lengend Snippet: ( a ) WT mice were injected i.p. with control IgG (isotype) or 50 μg of anti-CD3. The intestines were harvested 3 h post injection, and cDNA prepared from small sections of the indicated regions: qRT–PCR was used to measure the upregulation of ISGs as before. Error bars indicate s.e.m. of three mice per condition. Shown are the P values for anti-CD3 to isotype comparison, calculated by two-way analysis of variance. ( b , c ) The epithelial cell layer was harvested from small intestines of mice treated as in a and then sorted on a MACS column for CD45-positive (mainly IEL ( b )) and CD45-negative (mainly intestinal epithelial cells ( c )) fractions: qRT–PCR was performed as before. Expression is shown relative to TBP expression. Mean and s.e.m. of n =3 and the data shown are representative of four independent experiments. ( d ) Mice ( n =4) were injected i.p. with 25 μg anti-CD3 antibody or isotype, and after 3 h, the epithelial layer (EL) and LP layer (LPL) were extracted, and analysed by qRT–PCR as in b . ( e ) Mice were injected i.p. with 25 μg anti-CD3 antibody or isotype, and after 3 h the inguinal lymph nodes (LNs), spleen and IEL were isolated. TCRγδ + CD8αα + and TCRαβ + CD8αα + IEL were flow-sorted, and analysed by qRT–PCR, alongside sorted CD8 + cells from spleens and total LN cells. Normalized fold change from four experiments is shown. Statistical significance was determined by either unpaired t -tests ( b , c , e ) or using the Holm-Sidak method ( d ). * P <0.05, ** P <0.01, *** P <0.001. Flow cytometry of sorted cells and histology sections are in .

Article Snippet: Purified IEL and IEC fractions were either directly taken for flow cytometric analysis, sorting, or further purified by CD45 magnetic bead columns (Miltenyi).

Techniques: Injection, Control, Quantitative RT-PCR, Comparison, Expressing, Isolation, Flow Cytometry