Abstract
Purpose:
To identify, analyze and quantify stem cell marker positive cells in Human ALG from MMCR specimens. The ultimate goal is to develop cell based therapy for the treatment of dry eye disease from available MMCR specimens.
Methods:
A Tissue Microarray was created from 24 MMCR specimens to include the ALG. Immunofluorescence staining of ALG was performed using four stem cell markers ABCG2, CD90, CD133 and CD49f and were co-localized with Alpha Smooth Muscle Actin ASMA. A Semi automated image analysis was performed using the quantitative Vectra multispectral tissue imaging system and the inForm® software for acquisition of spectral data from each pixel in the image.
Results:
TMA demonstrated the presence of ALG in the majority of cores. Immunofluorescence staining of ALG showed positive stem cell marker staining of ABCG2 in lateral membranes of acinar cells, CD90 in association with ASMA positive myo-epithelial cells, CD49f with both acinar cells and myo-epithelial cells, CD133 was difficult to localize. The Vectra multispectral tissue imaging system was successful in identifying and quantifying stem cell markers positive cells in ALGs from Human MMCR specimens.
Conclusions:
MMCR specimens contain ALG that include stem cell marker positive cells co-localized to acinar or myo-epithelial cells. We attempted to identify stem cell marker positive cells as a first step to use of ALGs as a source of exocrine stem cell for cell based therapy. Localization of stem cell markers was in line with other studies. In addition multiple samples were tested from normal patients strengthening the finding of stem cell markers positive cell in normal human. Stem cell marker positive cells can be identified and further studies are needed to eventually develop a potential cell based treatment of severe dry eye disease.
Keywords: 526 eyelid •
576 lacrimal gland •
486 cornea: tears/tear film/dry eye