biostudies-literatureUnknown14(1)Sahoo A<h4>Background</h4>Mesenchymal stem cells (MSCs) have been proven to prevent and clear corneal scarring and limbal stem cell deficiency. However, using animal-derived serum in a culture medium raises the ethical and regulatory bar. This study aims to expand and characterize human limbus-derived stromal/mesenchymal stem cells (hLMSCs) for the first time in vitro in the xeno-free medium.<h4>Methods</h4>Limbal tissue was obtained from therapeutic grade corneoscleral rims and subjected to explant culture till tertiary passage in media with and without serum (STEM MACS XF; SM), to obtain pure hLMSCs. Population doubling time, cell proliferation, expression of phenotypic markers, tri-lineage differentiation, colony-forming potential and gene expression analysis were carried out to assess the retStem cell research & therapy89https://www.ebi.ac.uk/biostudies/studies/S-EPMC10105964biostudies-literatureExpansion and characterization of human limbus-derived stromal/mesenchymal stem cells in xeno-free medium for therapeutic applications.PMC10105964Sahoo ADamala MPrasad DBasu SJaffet JSingh VfalseExpansion and characterization of human limbus-derived stromal/mesenchymal stem cells in xeno-free medium for therapeutic applications.<h4>Background</h4>Mesenchymal stem cells (MSCs) have been proven to prevent and clear corneal scarring and limbal stem cell deficiency. However, using animal-derived serum in a culture medium raises the ethical and regulatory bar. This study aims to expand and characterize human limbus-derived stromal/mesenchymal stem cells (hLMSCs) for the first time in vitro in the xeno-free medium.<h4>Methods</h4>Limbal tissue was obtained from therapeutic grade corneoscleral rims and subjected to explant culture till tertiary passage in media with and without serum (STEM MACS XF; SM), to obtain pure hLMSCs. Population doubling time, cell proliferation, expression of phenotypic markers, tri-lineage differentiation, colony-forming potential and gene expression analysis were carried out to assess the ret2023-01-01T00:00:00Z2023 Apr2026-07-14T20:14:06.522Z2025-04-04T20:46:12.853ZS-EPMC101059643706173910.1186/s13287-023-03299-3