human bone marrow derived mscs c 12974  (PromoCell)

Journal: Investigative Ophthalmology & Visual Science

Article Title: Corneal Nerves Promote Alkali Burn Repair by Modulating Macrophages and Neutrophils via Calcitonin Gene-Related Peptide

doi: 10.1167/iovs.67.4.60

Figure Lengend Snippet: CGRP suppresses corneal tissue inflammation following alkali burn. ( A ) The volcano plot displayed the upregulated and downregulated genes in the corneas of mice from the CGRP-treated group compared with the PBS-treated group at 7 days post-injury ( n = 3). ( B ) The heatmap showed the differentially expressed genes in the corneas of mice from the CGRP-treated group compared with the PBS-treated group at 7 days post-injury ( n = 3). ( C ) The mRNA expression of the inflammatory factors IL-1β, iNOS, MCP-1, MMP3, and MMP9 was detected by qPCR on day 7 post-injury following CGRP treatment ( n = 4). ( D, E ) Representative flow cytometry plots and bar charts of CD45+ cells in the cornea after 7 days of treatment with CGRP following injury. ( F, G ) Representative micrographs and bar charts of macrophages in the cornea after 7 days treatment with topical CGRP or PBS ( n = 4). ( H ) Corneal macrophages were determined by flow cytometry ( n = 4). ( I, J ) Representative flow cytometry plots and bar charts of CD206+ macrophages in the cornea of the CGRP treatment group ( n = 4). ( K, L ) Representative micrographs and bar charts of neutrophils in the cornea after 7 days treatment with topical CGRP or PBS ( n = 4). (M) Corneal neutrophils were determined by flow cytometry ( n = 4). Data are shown as mean ± SD. ns: not significant; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: For the culture of neutrophils, the mouse bone marrow neutrophil isolation solution kit (Solarbio) was used to isolate neutrophils according to the manufacturer's instructions.

Techniques: Expressing, Flow Cytometry

Journal: Investigative Ophthalmology & Visual Science

Article Title: Corneal Nerves Promote Alkali Burn Repair by Modulating Macrophages and Neutrophils via Calcitonin Gene-Related Peptide

doi: 10.1167/iovs.67.4.60

Figure Lengend Snippet: Macrophage depletion results in delayed corneal repair after injury and abolishes the therapeutic effect of CGRP. ( A ) Experimental timeline showing the schedule of PLX5622 feeding, corneal alkali burn induction, topical CGRP or vehicle treatment, clinical evaluation, and tissue harvesting. ( B ) Representative images of corneal fluorescein staining in PLX5622-fed mice topically treated with CGRP or vehicle. ( C ) Percentage of injured corneal area in PLX5622-fed mice treated with CGRP or vehicle. Quantitatively analyzed by ImageJ software ( n = 6). ( D ) Representative images of cornea appearance in PLX5622-fed mice treated with CGRP or vehicle. ( E ) Quantification of the clinical score of corneal opacity ( n = 6). ( F, G ) Representative micrographs and quantification of TUNEL+ cells in corneas 7 days after alkali burn in mice fed with PLX5622 or control diet ( n = 4). ( H, I ) Representative micrographs and quantification of neutrophils in corneas 7 days after alkali burn in mice fed with PLX5622 or control diet ( n = 4). Data are shown as mean ± SD. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: For the culture of neutrophils, the mouse bone marrow neutrophil isolation solution kit (Solarbio) was used to isolate neutrophils according to the manufacturer's instructions.

Techniques: Staining, Software, TUNEL Assay, Control

Journal: Investigative Ophthalmology & Visual Science

Article Title: Corneal Nerves Promote Alkali Burn Repair by Modulating Macrophages and Neutrophils via Calcitonin Gene-Related Peptide

doi: 10.1167/iovs.67.4.60

Figure Lengend Snippet: Expression of the CGRP receptor in neutrophils and macrophages, and transcriptomic analysis of macrophages following CGRP treatment in vitro. ( A, B ) Light microscope image of bone marrow-derived macrophages and bone marrow-derived neutrophils ( n = 4). ( C ) CALCRL and RAMP1 expression was detected in bone marrow-derived macrophages and neutrophils using immunostaining. Green = CALCRL, red = RAMP1 ( n = 4). ( D ) Heatmap of selected significantly upregulated and downregulated genes depicting standardized gene expression values in CGRP-treated cells compared to PBS-treated cells. The colored circles next to the heatmap denote gene functions. ( E ) Volcano plot displaying upregulated and downregulated genes in macrophages from the CGRP-treated group versus the PBS-treated group. ( F ) GO enrichment analysis of significantly upregulated ( red ) and downregulated ( blue ) genes in CGRP-treated and saline-treated macrophages.

Article Snippet: For the culture of neutrophils, the mouse bone marrow neutrophil isolation solution kit (Solarbio) was used to isolate neutrophils according to the manufacturer's instructions.

Techniques: Expressing, In Vitro, Light Microscopy, Derivative Assay, Immunostaining, Gene Expression, Saline

Journal: Investigative Ophthalmology & Visual Science

Article Title: Corneal Nerves Promote Alkali Burn Repair by Modulating Macrophages and Neutrophils via Calcitonin Gene-Related Peptide

doi: 10.1167/iovs.67.4.60

Figure Lengend Snippet: CGRP enhances apoptotic, anti-inflammatory, and efferocytic functions of macrophages. ( A, E ) Representative micrographs and quantification of apoptosis in macrophages treated with CGRP in vitro ( n = 4). ( B, F ) Representative micrographs and quantification of apoptotic macrophages in the cornea following 7 days of topical CGRP treatment ( n = 4). ( C, G ) Representative micrographs and quantification of apoptosis in neutrophils treated with CGRP in vitro ( n = 4). ( D, H ) Representative micrographs and quantification of apoptotic neutrophils in the cornea following 7 days of topical CGRP treatment ( n = 4). ( I, J ) CGRP treatment promotes CD206 and Arg-1 mRNA expression in macrophages ( n = 4). ( K, L ) Representative micrographs and quantification of macrophage efferocytosis of apoptotic neutrophils ( n = 4). Data are shown as mean ± SD. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: For the culture of neutrophils, the mouse bone marrow neutrophil isolation solution kit (Solarbio) was used to isolate neutrophils according to the manufacturer's instructions.

Techniques: In Vitro, Expressing

Journal: Investigative Ophthalmology & Visual Science

Article Title: Corneal Nerves Promote Alkali Burn Repair by Modulating Macrophages and Neutrophils via Calcitonin Gene-Related Peptide

doi: 10.1167/iovs.67.4.60

Figure Lengend Snippet: Schematic illustration of corneal nerve-derived CGRP facilitating alkali burn repair through coordinated actions on neutrophils and macrophages.

Article Snippet: For the culture of neutrophils, the mouse bone marrow neutrophil isolation solution kit (Solarbio) was used to isolate neutrophils according to the manufacturer's instructions.

Techniques: Derivative Assay

Journal: bioRxiv

Article Title: TAR syndrome causal gene RBM8A is critical for embryonic bone development and proper Hedgehog signaling

doi: 10.64898/2026.04.21.718480

Figure Lengend Snippet: A. Immunofluorescence staining of cultured human bone marrow-derived mesenchymal stem cells showing CD90 (green) and RBM8A (red). Merged image (right) demonstrates RBM8A localization in CD90-positive cells. Scale bar = 50 µm. B. Whole-mount immunofluorescence staining of E10.5 mouse embryo section showing RBM8A (red), SOX9 (green), and merged image with DAPI (blue). RBM8A is broadly expressed, with high expression in the limb bud. In the neural crest, RBM8A expression is decreased in SOX9-positive regions. Scale bar = 200 µm. C. Whole-mount immunofluorescence staining of E14.5 mouse forelimb autopod sections showing RBM8A (red), SOX9 (green), and merged image with DAPI (blue). RBM8A is broadly distributed throughout the autopod, including SOX9-positive cartilage condensation. Scale bar = 200 µm. D. Whole-mount immunofluorescence staining of E14.5 mouse forelimb zeugopod longitudinal tissue section showing RBM8A (red), SOX9 (green), and merged image with DAPI (blue). RBM8A is detected in and around SOX9-positive skeletal condensations, including the central region of the developing cartilage element. Scale bar = 200 µm.

Article Snippet: Human bone marrow-derived MSCs were obtained from ATCC (catalog #: PCS-500-012) and cultured using Bone Marrow-Mesenchymal Stem Cell Basal Medium (ATCC, catalog #: PCS-500-030) with 7% FBS and growth factor supplements (15 ng/ml IGF-1, 125 ng/ml FGF2).

Techniques: Immunofluorescence, Staining, Cell Culture, Derivative Assay, Expressing

Journal: bioRxiv

Article Title: Plasma-Enabled Multiscale Coupling of Architecture and Biointerfaces Drives Osteogenesis in 3D-Printed Gyroid Scaffolds

doi: 10.64898/2026.04.16.718992

Figure Lengend Snippet: Fabrication and surface characterization of silver nanoparticle (AgNP)-functionalized PLA scaffolds. A) Schematic illustration of the scaffold fabrication and surface modification workflow: hexagonal honeycomb G-code was used to 3D-print PLA scaffolds, which were subsequently dip-coated in AgNO₃ solution, with or without prior incubation in polydopamine hydrochloride (PDA), followed by Plasma Electroless Reduction (PER) under H₂ gas to yield PLA+AgNP and PLA+PDA+AgNP constructs, respectively. B) Representative scanning electron microscopy (SEM) images of PLA+AgNP (top row) and PLA+PDA+AgNP (bottom row) scaffolds fabricated across a range of AgNO₃ concentrations (0–25 mM), with corresponding optical images of the scaffold surface shown as insets. Scale bars = 5 µm. C) SEM micrographs of L929 fibroblasts (top row) and human mesenchymal stem cells (hMSCs, bottom row) adhered to unmodified PLA HC, PLA HC+AgNP (0.7 mM AgNO₃), and PLA HC+PDA+AgNP (0.7 mM AgNO₃) scaffolds. Black arrows indicate representative cell–scaffold interactions. Scale bars = 15 µm.

Article Snippet: The cells were incubated in mesenchymal stem cell complete media (Basal media: ATCC, USA: Cat no: PCS-500-030 and growth kit: ATCC, USA: Cat no: PCS-500-041) for five days.

Techniques: Modification, Incubation, Clinical Proteomics, Construct, Electron Microscopy