Purchase this article with an account.
Shubha Tiwari, Mohammad Ali, Murali Mohan Sagar Balla, Milind Naik, Santosh Honavar, Vijay Anand Palkonda, Geeta Vemuganti; Stem-like Cells in Serum Free In-vitro Cultures of Human Lacrimal Gland. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2191.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Tear film deficiency due to lacrimal gland dysfunction or damage is an important cause of ocular morbidity. Restoration of gland function by transplantation of autologous ex-vivo expanded stem cells located in the lacrimal gland is one of the options that could relieve this problem. We have previously reported the presence of stem-like and functionally competent differentiated cells in in-vitro cultures of human lacrimal gland. The present study focuses on the formation of ‘lacrispheres’ under serum-free condition and the expression of stemness in them.
Fresh human lacrimal gland tissues (n=7) from patients undergoing exenteration were harvested for cultures after IRB approval. The gland was processed by enzymatic digestion using a cocktail of collagenase and hyaluronidase. The isolated cells were plated on ultralow attachment plates as group of two-three cells and fed with HepatoStim supplemented with epidermal growth factor, fibroblast growth factor and N2. The spheres were pulse-labeled with BrDU, analyzed for the expression of CD117 by immunocytochemistry and their colony forming efficiency was assessed on Matrigel.
Serum free cultures demonstrate spheres from human lacrimal gland in-vitro within 2-3 days of plating. These spheres grow in size over 3-14 days and can be serially passaged to generate secondary spheres. Anti-BrDU labeling of these spheres indicate the presence of 3.8% of high intensity cells at the periphery and about 3% dull intensity cells at the center. These also show positive labeling for CD117 and formation of clones in Matrigel (CFU 3.1%) indicating the presence of stemness.
This is the first report on the generation of ‘lacrisphers’ in human lacrimal gland cultures. It strengthens our initial reports that human lacrimal gland has a storehouse of stem cells that can be maintained in-vitro and could possibly serve as potential source of cell therapy for the regeneration of the functionally compromised gland.
This PDF is available to Subscribers Only