Panel of fluorescent-tagged monoclonal antibodies and their immune-cell subtype targets.
| Group | Antibodies | Cell types |
|---|---|---|
| I | CD3 (FITC) + CD45RA (RPE) | B Lymphocytes, T Lymphocytes |
| II | CD11b (A488) + CD161 (RPE) | Natural Killer Cells, Macrophages + Monocytes + Dendritic Cells + Granulocytes |
| III | CD4 (A488) + CD8a (RPE) | Cytotoxic T-Cells + Natural Killer Cells, Helper T-Cells + Monocytes |
| IV | RT1B (RPE) + IgM (FITC) | B Lymphocytes |
| V | CD11b (Pacific Blue) + RT1B (FITC) + CD86 (RPE) | Macrophages + Monocytes + Dendritic Cells + Granulocytes, Dendritic Cells + Macrophages + B Lymphocytes |
| VI | CD68 (A488) + CD11b (Pacific Blue) + CD163 (RPE) | Macrophages, Macrophages + Monocytes + Dendritic Cells + Granulocytes |
| VII | CD43 (FITC) | Leukocytes |
| VIII | Unstained control | N/A |
| IX | CD45 (A488) | Leukocytes + B Lymphocytes |
Samples of rat peripheral blood were divided and stained with fluorescent-tagged monoclonal antibodies. Each antibody targeted a specific protein associated with an immune-cell subtype. The mean fluorescence was quantified as a percentage of the total of 40,000 cells per sample.
The authors acknowledge the assistance of Xiaocun Sun in running the statistical analysis. The authors acknowledge the assistance of Robert Donnell for analysis of the H&E-stained histological slides. The authors acknowledge the assistance of Lisa Amelse in performing the antibody staining. The authors acknowledge the assistance of Stacey Stephenson in performing the sciatic nerve defect surgery. The authors acknowledge the assistance of Tom Masi in seeding the MSCs on the scaffolds. The authors acknowledge the assistance of Angela Kites in figure visualization. Dr. Mohamed Abouelkhair is now affiliated with Rowan University’s Shreiber School of Veterinary Medicine (Department of Diagnostic Medicine and Pathobiology, Glassboro, NJ 08028, USA) following the completion of this research, which was conducted while at the University of Tennessee. Briana Lewis is now affiliated with Mississippi State University (Department of Animal Science, Mississippi State, MS 39762, USA) following the completion of this research, which was conducted while at the University of Tennessee.
MEHT: Conceptualization, Data curation, Formal analysis, Investigation, Project administration, Visualization, Writing—original draft, Writing—review & editing. MAA: Methodology, Resources, Software, Supervision, Validation, Writing—review & editing. SDN: Investigation, Methodology, Writing—review & editing. BL: Investigation, Writing—review & editing. DEA: Conceptualization, Funding acquisition, Supervision, Writing—review & editing. MD: Conceptualization, Data curation, Funding acquisition, Project administration, Resources, Supervision, Validation, Writing—review & editing. All authors consent to publication.
The authors declare that there are no conflicts of interest.
The study complied with the 2024 Helsinki Declaration and “Guide for the Care and Use of Laboratory Animals”. All procedures were conducted in accordance with PHS guidelines for the humane treatment of animals under approved protocols established through the University of Tennessee’s Institutional Animal Care and Use Committee (IACUC) (IACUC# 2574-0921). The human adipose-derived mesenchymal stem cells used for the experiments described in this manuscript were derived from adipose tissue collected from patients undergoing pannulectomies at the University of Tennessee Medical Center. The collections were performed under an approved Institutional Review Board (IRB) protocol (IRB#3995). Written informed consent was obtained from each patient, per the IRB protocol.
Patient consent was obtained in accordance with the IRB-approved protocol at the University of Tennessee Medical Center.
Not applicable.
The raw data supporting the conclusions of this manuscript will be made available by the authors, without undue reservation, to any qualified researcher.
This article was funded by the University of Tennessee: Office of Research, Innovation, and Economic Development Seed Award. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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