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The Role of Nerve Growth Factor in Maintaining Proliferative Capacity, Colony‐Forming Efficiency, and the Limbal Stem Cell Phenotype

Lookup NU author(s): Dr Sai Kolli, Dr Sanja Bojic, Dr Ali Ghareeb, Dr Marzena Kurzawa-Akanbi, Professor Francisco Figueiredo, Professor Majlinda Lako

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

Nerve growth factor (NGF) has demonstrated great benefit in the treatment of neurotrophic corneal ulcers. There is evidence for multiple modes of action in promoting corneal healing, but only indirect evidence exists for NGF’s effects on limbal stem cells (LSCs). Understanding the role of NGF in LSC biology will improve our understanding of paracrine regulation of the limbal niche and the design of stem cell-based therapies for conditions such as limbal stem cell deficiency. In this manuscript, we studied the regulation of NGF signalling components during LSC differentiation and the role of NGF in LSC proliferation and maintenance of the stem cell phenotype. LSC differentiation was induced by prolonged (40 day) culture which resulted in a significant increase in cell size, decrease in colony-forming efficiency and expression of putative LSC markers. A protein microarray measuring expression of 248 signalling proteins indicated the low affinity NGF receptor p75NTR to be the most downregulated protein upon differentiation. Further confirmation by Western blotting and RT-qPCR indicated that NGF and p75NTR are expressed in early LSC cultures and downregulated upon differentiation. LSC cultures grown in the presence of anti-NGF antibody showed decreased colony-forming efficiency, DNA replication and expression of putative LSC markers ABCG2 and C/EBPd. Supplementation of LSC culture medium with NGF extended the lifespan of LSC cultures in vitro and increased the expression of putative LSC markers ΔNp63α and ABCG2. Taken together our data indicate that NGF signalling is a key promotor of LSC proliferation, colony forming-efficiency and a maintainer of the LSC phenotype.


Publication metadata

Author(s): Kolli S, Bojic S, Ghareeb AE, Kurzawa-Akanbi M, Figueiredo FC, Lako M

Publication type: Article

Publication status: Published

Journal: Stem Cells

Year: 2019

Volume: 37

Issue: 1

Print publication date: 01/01/2019

Online publication date: 31/12/2018

Acceptance date: 28/08/2018

Date deposited: 05/09/2018

ISSN (print): 1066-5099

ISSN (electronic): 1549-4918

Publisher: Wiley

URL: https://doi.org/10.1002/stem.2921

DOI: 10.1002/stem.2921


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