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Updated: Aug 11, 2026
Murine Dermal Fibroblast Isolation by FACS
Published on: January 7, 2016
A fibroblast-associated antigen: characterization in fibroblasts and immunoreactivity in smooth muscle differentiated
L Rønnov-Jessen1, J E Celis, B Van Deurs
1Department of Anatomy, Panum Institute, Copenhagen, Denmark.
Insights
A novel fibroblast-associated antigen, recognized by monoclonal antibody 1B10, helps distinguish smooth muscle-differentiated fibroblasts from vascular smooth muscle cells. This antigen is found on the cell surface and within vesicular compartments.
Area of Science:
- Cell Biology
- Immunology
- Histology
Background:
- Fibroblasts exhibiting smooth muscle differentiation are common in human breast tissue.
- Distinguishing these differentiated fibroblasts from vascular smooth muscle cells is crucial for accurate tissue analysis.
Purpose of the Study:
- To characterize a fibroblast-associated antigen using monoclonal antibody (MAb) 1B10.
- To determine if MAb 1B10 can differentiate smooth muscle-differentiated fibroblasts from vascular smooth muscle cells.
Main Methods:
- Immunofluorescence cytochemistry was employed to detect the antigen.
- Ultrastructural analysis examined the antigen's localization within cellular compartments.
- Western blot analysis identified the molecular weight of recognized proteins.
Main Results:
- The antigen was detected on the fibroblast cell surface and in vesicular compartments (endosomes, lysosomes).
- Ultrastructural analysis revealed the antigen in coated pits and lysosomal structures.
- MAb 1B10 recognized multiple protein bands, with specific associations to the cell membrane and lysosomes.
- 1B10 immunoreactivity was specific to fibroblasts and smooth muscle-differentiated fibroblasts, differentiating them from vascular smooth muscle cells.
Conclusions:
- Monoclonal antibody 1B10 identifies a specific antigen on smooth muscle-differentiated fibroblasts.
- This antigen serves as a reliable marker for distinguishing these cells from vascular smooth muscle cells.
Abstract:
Fibroblasts with smooth muscle differentiation are frequently derived from human breast tissue. Immunofluorescence cytochemistry of a fibroblast-associated antigen recognized by a monoclonal antibody (MAb), 1B10, was analyzed with a view to discriminating smooth muscle differentiated fibroblasts from vascular smooth muscle cells. The antigen was detected on the cell surface and in cathepsin D-positive and acridine orange-accumulating vesicular compartments of fibroblasts. Ultrastructurally, the antigen was revealed in coated pits and in endosomal and lysosomal structures. 1B10 recognized three major brands migrating at apparent Mr of 38,000, 45,000, and 80,000, in addition to many minor bands between Mr 45,000 and 97,000, including Mr 52,000. The Mr 45,000 and 38,000 were associated with the cell membrane and Mr 52,000 as well as Mr 38,000 were associated with the lysosomes. The 1B10 immunoreactivity was specific to fibroblasts and smooth muscle differentiated fibroblasts within the context of vascular smooth muscle cells.
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