Evaluation of methods for cultivating limbal mesenchymal stromal cells

Cytotherapy
Laura J BrayDamien G Harkin

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

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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
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Apr 15, 2014·Cytotherapy·Shahla Hamza Al-SaqiCecilia Götherström
Aug 15, 2014·Cell and Tissue Banking·Shahla Hamza Al-SaqiCecilia Götherström
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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 16, 2020·Investigative Ophthalmology & Visual Science·Yu-Ting XiaoXin-Yue Zhao

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