BSc Human Biology at Universitat Pompeu Fabra, Barcelona (Spain) MSc Molecular Bioengineering at Biotec, Technisches Universität Dresden (Germany) Currently a PhD student at Trinity College Dublin developing therapies for corneal transplantation using Tissue Engineering approaches.
Jan 2014 - Aug 2015
- Max Bergmann Center of Biomaterials (MBC)
- Dresden, Germany
- Master's Student
- Master thesis: "Development of a three dimensional bioengineered construct of the human corneal limbus"
Research Items (6)
- May 2019
The global shortage of donor corneas for transplantation has led to corneal bioengineering being investigated as a method to generate transplantable tissues. Decellularized corneas are among the most promising materials for engineering corneal tissue since they replicate the complex structure and composition of real corneas. Decellularization is a process that aims to remove cells from organs or tissues resulting in a cell-free scaffold consisting of the tissues extracellular matrix. Here different decellularization techniques are described, including physical, chemical and biological methods. Analytical techniques to confirm decellularization efficiency are also discussed. Different cell sources for the recellularization of the three layers of the cornea, recellularization methods used in the literature and techniques used to assess the outcome of the implantation of such scaffolds are examined. Studies involving the application of decellularized corneas in animal models and human clinical studies are discussed. Finally, challenges for this technology are explored involving scalability, automatization and regulatory affairs
Purpose: To identify biochemical cues that could promote a keratocyte-like phenotype in human corneal stromal cells that had become fibroblastic when expanded in serum-supplemented media while also examining the effect on cell proliferation and migration. Methods: Proliferation was assessed by PrestoBlueTM, morphology was monitored by phase contrast microscopy, phenotype was analysed by real-time polymerase chain reaction (qPCR), immunochemistry and flow cytometry, and migration was studied with a scratch assay. Results: Ascorbic Acid (AA), Retinoic Acid (RA), Insulin-Transferrin-Selenium (ITS), Insulin-like Growth Factor 1 (IGF-1) and 3-isobutyl-1-methylxanthine (IBMX) promoted a dendritic morphology, increased the expression of keratocyte markers, such as keratocan, aldehyde dehydrogenase 3 family member A1 (ALDH3A1) and CD34, and prevented myofibroblast differentiation, while in some cases increasing proliferation. Transforming Growth Factor beta 1 (TGF-β1) and 3 (TGF-β3) promoted the differentiation towards myofibroblasts, with increased expression of α-SMA. Fibroblast Growth Factor 2 (FGF-2) supported a fibroblastic phenotype while Platelet-Derived Growth Factor Homodimer B (PDGF-BB) induced a pro-migratory fibroblastic phenotype. A combination of all the pro-keratocyte factors was also compared to the serum-free only, which significantly increased CD34 and keratocan expression. Conclusions: Partially recovery towards a quiescent keratocyte-like phenotype was achieved by the removal of serum and the addition of AA, IGF-1, RA, ITS and IBMX to a basal medium. These findings can be used to develop cell-based corneal therapies and to study corneal diseases in vitro.
Limbal stromal cells (LSCs) from the human ocular surface display mesenchymal stromal cell characteristics in vitro. In this study, we isolated cells from the porcine limbal stroma (pLSCs), characterised them, and evaluated their ability to support angiogenesis and the culture of porcine limbal epithelial stem cells (pLESCs). The isolated cells adhered to plastic and grew in monolayers in vitro using serum-supplemented or serum-free medium. The pLSCs demonstrated expression of CD29, and cross-reactivity with anti-human CD45, CD90, CD105, CD146, and HLA-ABC. However, expression of CD105, CD146 and HLA-ABC reduced when cultured in serum-free medium. PLSCs did not undergo adipogenic or osteogenic differentiation, but differentiated towards the chondrogenic lineage. Isolated cells were also co-cultured with human umbilical vein endothelial cells (HUVECs) in star-shaped Poly(ethylene glycol) (starPEG)-heparin hydrogels to assess their pericyte capacity which supported angiogenesis networks of HUVECs. PLSCs supported the three dimensional HUVEC network for 7 days. The isolated cells were further growth-arrested and evaluated as feeder cells for pLESC expansion on silk fibroin membranes, as a potential carrier material for transplantation. PLSCs supported the growth of pLESCs comparably to murine 3T3 cells. In conclusion, although pLSCs were not completely comparable to their human counterpart, they display several mesenchymal-like characteristics in vitro.
- Oct 2016
Purpose With the increasing demand for corneal replacements and a shortage of suitable donors, tissue engineering and cell based therapies have emerged as a possible solution. Understanding the influence that biochemical factors have on corneal wound healing is needed to improve tissue regeneration. Our aim was to investigate the effect of several molecules on human corneal stromal cells. Methods Corneal fibroblasts were serum-starved for 72 h in DMEM/F12 and then exposed to specific reagents for 7–14 days. The reagents examined in this study were ascorbic acid (AA), retinoic acid (RA), FGF-2, PDGF-BB, IGF, TGF-β1, TGF-β3, IBMX and insulin-transferrin-selenium (ITS). Proliferation was assessed by AlamarBlue assay, general morphology was monitored by bright field microscopy and the expression of keratocan, ALDH3A1 and smooth muscle actin (α-SMA) at the mRNA level was quantified by qPCR. Immunohistochemistry and western blotting were also performed to assess protein expression. Results AA, IGF-1, RA and ITS increased the expression of keratocyte markers such as keratocan and ALDH3A1, while not increasing fibrotic/myofibroblast marker α-SMA. FGF-2 and PDGF-BB decreased expression of keratocan and ALDH3A1 without up-regulating α-SMA, suggesting a fibroblastic phenotype. TGF-β1 and TGF-β3 were shown to up-regulate α-SMA expression but TGF-β3 also up-regulated keratocan. All reagents apart from AA resulted in an increase in cell proliferation by day 7. Conclusions This study demonstrates the influence different biochemical cues have on corneal stromal cells. These findings will be useful in developing the next generation of therapies for corneal diseases and injuries.
Poster presented at Bioengineering in Ireland 22 as part of the ”Early Researchers” section
Awards & Achievements (3)
Award · Sep 2018
ISER Travel Fellowship
Scholarship · May 2017
The Network of Excellence in Corneal Regeneration. COST Action BM1302 (ECOST-STSM-BM1302-010517-088265)
Scholarship · Sep 2014
Deutschland Stipendium (sponsored by BASF)