Human umbilical cord mesenchymal stem cells improve generation of spheres in human corneal epithelial cells and promotes cell migration

Abstract

Event Abstract Back to Event Human umbilical cord mesenchymal stem cells improve generation of spheres in human corneal epithelial cells and promotes cell migration Siti M. Azmi1*, Nur Zawani M. Kamal1, Wai H. Ng1, Rafeezul Mohamed1, Jun J. Tan1 and Bakiah Shaharuddin1 1 Universiti Sains Malaysia, Regenerative Medicine Cluster, Advanced Medical and Dental Institute, Malaysia Transplantation of cornea limbal stem cells is the contemporary cellular therapy for corneal surface regeneration. Cells in three-dimensional (3D) cultures mimic the microenvironment in vivo thus improve the engraftment and survival rates of cellular transplantation (Fennema, Rivron et al. 2013, Kawaguchi, Machida et al. 2013, Oltolina, Zamperone et al. 2015, Wan, Wang et al. 2015). Umbilical cord-derived mesenchymal stem cells (UCMSCs) possess regenerative capability and are rich with paracrine factors (Guo, Sun et al. 2015). The aim of this project was to study human cornea epithelial cells (hCEC) sphere formation under the influence of UCMSC, and its effect on cellular migration, and to investigate the stem cell markers expression in 3D co-cultures of hCEC-USMSC. In this study, spheres were generated by hanging drop technique from either hCEC alone, or combination with UCMSC using sphere induction medium (50% DMEM and 50% DKSFM supplemented with 10 ng/ml EGF and 20 ng/ml bFGF). Spontaneous sphere formation was performed on low cell-attachment plate and served as control. Spheres were harvested on day 4 and characterised. Cellular migration was evaluated by scratch migration assay in sphere cultures versus control. The results showed that hCEC sphere formation was improved in 3D by the hanging drop method compared to spontaneous generation on low adherent surface. hCEC sphere formation improved by the presence of UCMSC compared to hCEC alone (Fig 1). Immunofluorescence revealed that hCEC in spheres maintained their epithelial stem cell marker p63 without expression of differentiation marker cytokeratin 3 (Fig 2). Preliminary study on cell migration demonstrated capability for gap closure was enhanced in sphere cultures. In conclusion, UCMSC improves generation of sphere in HCECs. 3D sphere cultures showed a better condition for maintaining epithelial stem cell markers. Further functional studies to examine the effects of spherical hCEC-UCMSC co-culture for corneal regeneration will be investigated further. Figure 1 Figure 2 Acknowledgements This work was funded by Fundamental Research Grant Scheme (203.CIPPT.6711508) Ministry of Higher Education Malaysia. Keywords: 3D culture, Umbilical cord mesenchymal stem cell, Corneal epithelial, spherical culture, Cord Factors Conference: 6th Malaysian Tissue Engineering and Regenerative Medicine Scientific Meeting (6th MTERMS) 2016 and 2nd Malaysian Stem Cell Meeting, Seberang Jaya, Penang, Malaysia, 17 Nov - 18 Nov, 2016. Presentation Type: Poster Topic: Stem cells and Regenerative Medicine Citation: Azmi SM, Kamal NM, Ng WH, Mohamed R, Tan JJ and Shaharuddin B (2016). Human umbilical cord mesenchymal stem cells improve generation of spheres in human corneal epithelial cells and promotes cell migration. Front. Bioeng. Biotechnol. Conference Abstract: 6th Malaysian Tissue Engineering and Regenerative Medicine Scientific Meeting (6th MTERMS) 2016 and 2nd Malaysian Stem Cell Meeting. doi: 10.3389/conf.FBIOE.2016.02.00019 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 08 Dec 2016; Published Online: 19 Dec 2016. * Correspondence: Ms. Siti M Azmi, Universiti Sains Malaysia, Regenerative Medicine Cluster, Advanced Medical and Dental Institute, Kepala Batas, Pulau Pinang, 13200, Malaysia, maisura@usm.my Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Siti M Azmi Nur Zawani M Kamal Wai H Ng Rafeezul Mohamed Jun J Tan Bakiah Shaharuddin Google Siti M Azmi Nur Zawani M Kamal Wai H Ng Rafeezul Mohamed Jun J Tan Bakiah Shaharuddin Google Scholar Siti M Azmi Nur Zawani M Kamal Wai H Ng Rafeezul Mohamed Jun J Tan Bakiah Shaharuddin PubMed Siti M Azmi Nur Zawani M Kamal Wai H Ng Rafeezul Mohamed Jun J Tan Bakiah Shaharuddin Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.