biostudies-literatureMei HNEI NIH HHSNational Eye InstituteResearch to Prevent BlindnessCalifornia Institute for Regenerative MedicineNational Eye Institute grantse0186238https://www.ebi.ac.uk/biostudies/studies/S-EPMC5636133biostudies-literatureUnknown12(10)The most efficient method to expand limbal stem cells (LSCs) in vitro for clinical transplantation is to culture single LSCs directly on growth-arrested mouse fibroblast 3T3 cells. To reduce possible xenobiotic contamination from 3T3s, primary human adipose-derived stem cells (ASCs) were examined as feeder cells to support the expansion of LSCs in vitro. To optimize the ASC-supported culture, freshly isolated limbal epithelial cells in the form of single cells (SC-ASC) or cell clusters (CC-ASC) were cultured using three different methods: LSCs seeded directly on feeder cells, a 3-dimensional (3D) culture system and a 3D culture system with fibrin (fibrin 3D). The expanded LSCs were examined at the end of a 2-week culture. The standard 3T3 culture served as control. Expansion of SC-ASC showPloS oneHuman adipose-derived stem cells support the growth of limbal stem/progenitor cells.PMC56361331R01EY021797R01 EY0217975P30EY000331P30 EY000331TR2-01768Nakatsu MNMei HDeng SXChen FVGonzalez SBaclagon ERfalseHuman adipose-derived stem cells support the growth of limbal stem/progenitor cells.The most efficient method to expand limbal stem cells (LSCs) in vitro for clinical transplantation is to culture single LSCs directly on growth-arrested mouse fibroblast 3T3 cells. To reduce possible xenobiotic contamination from 3T3s, primary human adipose-derived stem cells (ASCs) were examined as feeder cells to support the expansion of LSCs in vitro. To optimize the ASC-supported culture, freshly isolated limbal epithelial cells in the form of single cells (SC-ASC) or cell clusters (CC-ASC) were cultured using three different methods: LSCs seeded directly on feeder cells, a 3-dimensional (3D) culture system and a 3D culture system with fibrin (fibrin 3D). The expanded LSCs were examined at the end of a 2-week culture. The standard 3T3 culture served as control. Expansion of SC-ASC show2017-01-01T00:00:00Z20172025-04-26T20:05:46.429Z2019-03-27T02:58:41ZS-EPMC56361332902011910.1371/journal.pone.0186238