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Sheyla Gonzalez, Hua Mei, Martin N Nakatsu, Elfren Ray Baclagon, Sophie Xiaohui Deng; Human Bone Marrow-Derived Mesenchymal Stem Cells Support the Growth of Limbal Epithelial Progenitor/Stem Cells. Invest. Ophthalmol. Vis. Sci. 2014;55(13):522. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To test whether human bone marrow-derived mesenchymal stem cells (BM-MSCs) can serve as feeder cells to expand human limbal epithelial progenitor/stem cells (LSCs).
Primary human LSCs were isolated and seeded as single cells and cell clusters in the presence of BM-MSCs. LSCs were cultured directly on a BM-MSC monolayer (standard method), in a 3-dimensional (3-D) method, and 3-D fibrin method. Single LSCs cultured on a 3T3-J2 monolayer were used as a control. Cultured LSCs were analyzed for cell morphology, cell growth rate, and expression level of putative stem cell markers.
LSCs did not grow efficiently using the direct or standard method. LSCs in the cell cluster form showed a 2.5-fold higher cell growth rate using the 3-D method than in the control. LSCs cultured by 3-D and 3-D fibrin methods had a similar K14 mRNA expression level and a significantly lower K12 mRNA expression compared to the control (decreased by 34% and 95%, respectively; p<0.05). There was no difference in the percentage of K14+ and K12+ cells among all culture methods. The percentage of p63α bright cells in the 3-D (13.6%) and 3-D fibrin method (11%) was comparable to that in the control (12.8%, p>0.05).
BM-MSCs could support the growth and maintenance of LSCs. The expansion rate was the highest in the novel 3-D culture method.
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