title: A 3D culture system enhances the ability of human bone marrow stromal cells to support the growth of limbal stem/progenitor cells.
abstract: The standard method of cultivating limbal epithelial progenitor/stem cells (LSCs) on a monolayer of mouse 3T3 feeder cells possesses the risk of cross-contamination in clinical applications. Human feeder cells have been used to eliminate this risk; however, efficiency from xenobiotic-free cultures on a monolayer appears to be lower than in the standard method using 3T3 cells. We investigated whether bone marrow stromal cells (BMSCs), also known as bone marrow-derived mesenchymal stem cells, could serve as feeder cells for the expansion of LSCs in the 3-dimensional (3D) system. Primary single human LSCs on a monolayer of 3T3s served as the control. Very poor growth was observed when single LSCs were cultured on BMSCs. When LSC clusters were cultured on a BMSC monolayer (CC-BM), 3D culture system (3D CC-BM) and fibrin 3D system (fibrin 3D CC-BM), the 3D CC-BM method supported a greater LSC expansion. The 3D CC-BM system produced a 2.5-fold higher cell growth rate than the control (p<0.05). The proportion of K14(+) and p63α(bright) cells was comparable to those in the control (p>0.05), whereas the proportion of K12(+) cells was lower (p<0.05). These results indicate that BMSCs can efficiently support the expansion of the LSC population in the 3D culture.
authors.affiliation: Cornea Division, Stein Eye Institute, University of California, Los Angeles, CA, USA.
authors.forename: Sheyla
authors.initials: S
authors.lastname: González
authors.affiliation: Cornea Division, Stein Eye Institute, University of California, Los Angeles, CA, USA.
authors.forename: Hua
authors.initials: H
authors.lastname: Mei
authors.affiliation: Cornea Division, Stein Eye Institute, University of California, Los Angeles, CA, USA.
authors.forename: Martin N
authors.initials: MN
authors.lastname: Nakatsu
authors.affiliation: Cornea Division, Stein Eye Institute, University of California, Los Angeles, CA, USA.
authors.forename: Elfren R
authors.initials: ER
authors.lastname: Baclagon
authors.affiliation: Cornea Division, Stein Eye Institute, University of California, Los Angeles, CA, USA. Electronic address: deng@jsei.ucla.edu.
authors.forename: Sophie X
authors.initials: SX
authors.lastname: Deng
mesh_terms: D016475:3T3 Cells; D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology*; D015820:Cadherins / Q000378:metabolism; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology; D061252:Feeder Cells; D006801:Humans; D007150:Immunohistochemistry; D017493:Leukocyte Common Antigens / Q000378:metabolism; D016850:Limbus Corneae / Q000166:cytology*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008853:Microscopy; D010641:Phenotype; D060888:Real-Time Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D018952:Antigens, CD34; D015820:Cadherins; D017493:Leukocyte Common Antigens
keywords: Bone marrow derived-mesenchymal stem cell; Bone marrow stromal cells; Corneal epithelium; Limbal stem cell deficiency; Limbal stem cells