Abstract
Mesenchymal stromal cells (MSC) with similar properties to bone marrow-derived mesenchymal stromal cells (BM-MSC) have recently been grown from the limbus of the human cornea. We have evaluated methods for culturing human limbal MSC (L-MSC). Four basic strategies were compared: serum-supplemented medium (10% fetal bovine serum; FBS), standard serum-free medium supplemented with B-27, epidermal growth factor and fibroblast growth factor 2, or one of two commercial serum-free media, defined keratinocyte serum-free medium (Invitrogen) and MesenCult-XF® (Stem Cell Technologies). The resulting cultures were examined using photography, flow cytometry (for CD34, CD45, CD73, CD90, CD105, CD141 and CD271), immunocytochemistry (alpha-smooth muscle actin; α-sma), differentiation assays (osteogenesis, adipogenesis and chrondrogenesis) and co-culture experiments with human limbal epithelial (HLE) cells. While all techniques supported the establishment of cultures to varying degrees, sustained growth and serial propagation were only achieved in 10% FBS medium or MesenCult-XF medium. Cultures established in 10% FBS medium were 70-80% CD34(-) CD45(-) CD90 (+) CD73 (+) CD105 (+) , approximately 25% α-sma (+) and displayed multipotency. Cultures...Continue Reading
References
Jul 1, 1986·The Journal of Cell Biology·A SchermerT T Sun
Apr 1, 1994·The Journal of Clinical Investigation·T J RaifeS R Lentz
Apr 5, 1997·Lancet·G PellegriniM De Luca
Nov 14, 1998·Lupus·M C Boffa, M Karmochkine
Jul 13, 2000·The New England Journal of Medicine·R J TsaiJ K Chen
Sep 6, 2001·Ophthalmology·N KoizumiS Kinoshita
Jun 5, 2002·Ophthalmology·Paolo TotiAldo Caporossi
Aug 3, 2002·Ophthalmology·Martin GrueterichScheffer C G Tseng
Oct 28, 2003·Investigative Ophthalmology & Visual Science·Annie JosephHarminder S Dua
Aug 20, 2004·The British Journal of Ophthalmology·D G HarkinA J G Apel
Jul 30, 2005·Stem Cells·Yiqin DuJames L Funderburgh
Aug 23, 2006·Cytotherapy·M DominiciEm Horwitz
Apr 28, 2007·Cytotherapy·P-F ChoongK-Y Then
Nov 21, 2007·Seminars in Cell & Developmental Biology·Gary BrookeKerry Atkinson
Jan 13, 2009·Investigative Ophthalmology & Visual Science·Masahiro OmotoKazuo Tsubota
Feb 21, 2009·Journal of Tissue Engineering and Regenerative Medicine·Rakhi PalSatish Totey
Jun 25, 2010·The New England Journal of Medicine·Paolo RamaGraziella Pellegrini
Oct 26, 2010·Experimental Eye Research·S Louise AinscoughDamien G Harkin
Citations
Jun 27, 2013·Stem Cell Research & Therapy·Khurram HashmaniHarminder Singh Dua
Jun 18, 2014·In Vitro Cellular & Developmental Biology. Animal·Ying ZhouHuiyong Zhu
Dec 12, 2012·Current Opinion in Organ Transplantation·Richard Verbeek
Oct 8, 2013·Cytotherapy·Laura J BrayDamien G Harkin
Apr 27, 2013·Cell Biology International·Jinny Jung YoonTrevor Sherwin
Nov 18, 2014·Stem Cells·Damien G HarkinBrendan G Cronin
Oct 15, 2013·Cell Proliferation·S GottipamulaR N Seetharam
Oct 12, 2015·Cytotherapy·Laura E SidneyAndrew Hopkinson
Apr 15, 2014·Cytotherapy·Shahla Hamza Al-SaqiCecilia Götherström
Jul 4, 2015·Cytotherapy·Kah Yong TanSteve Kw Oh
Aug 15, 2014·Cell and Tissue Banking·Shahla Hamza Al-SaqiCecilia Götherström
Dec 17, 2014·Experimental Eye Research·Peter J GilliesDamien G Harkin
Mar 24, 2015·PloS One·Rishi Man ChughLakshmana Kumar Yerneni
Jun 15, 2016·Stem Cell Research & Therapy·Laura TomaselloCarla Giordano
Jan 1, 2013·Journal of Functional Biomaterials·Samantha L WilsonAndrew Hopkinson
Dec 8, 2017·Stem Cells International·Richárd NagymihályGoran Petrovski
Sep 27, 2014·World Journal of Stem Cells·Jinny J YoonTrevor Sherwin
Jan 18, 2018·Cell Transplantation·Damien G HarkinNadia Zakaria
Jun 25, 2020·International Journal of Molecular Sciences·Sebastian G WalterFrank A Schildberg
May 23, 2017·Journal of the Chinese Medical Association : JCMA·Shu-I PaoChang-Min Liang
Jul 27, 2017·Scientific Reports·Julia Fernández-PérezLaura J Bray
Dec 6, 2017·Cytotechnology·Mehmet Gürdalİsmet Durak
Dec 16, 2020·Investigative Ophthalmology & Visual Science·Yu-Ting XiaoXin-Yue Zhao