7 research outputs found
Human conjunctiva organoids to study ocular surface homeostasis and disease
The conjunctival epithelium covering the eye contains two main cell types: mucus-producing goblet cells and water-secreting keratinocytes, which present mucins on their apical surface. Here, we describe long-term expanding organoids and air-liquid interface representing mouse and human conjunctiva. A single-cell RNA expression atlas of primary and cultured human conjunctiva reveals that keratinocytes express multiple antimicrobial peptides and identifies conjunctival tuft cells. IL-4/-13 exposure increases goblet and tuft cell differentiation and drastically modifies the conjunctiva secretome. Human NGFR+ basal cells are identified as bipotent conjunctiva stem cells. Conjunctival cultures can be infected by herpes simplex virus 1 (HSV1), human adenovirus 8 (hAdV8), and SARS-CoV-2. HSV1 infection was reversed by acyclovir addition, whereas hAdV8 infection, which lacks an approved drug therapy, was inhibited by cidofovir. We document transcriptional programs induced by HSV1 and hAdV8. Finally, conjunctival organoids can be transplanted. Together, human conjunctiva organoid cultures enable the study of conjunctival (patho)-physiology.</p
Optimization of Media Change Intervals through Hydrogels Using Mathematical Models
Three-dimensional cell culture in engineered hydrogels
is increasingly
used in tissue engineering and regenerative medicine. The transfer
of nutrients, gases, and waste materials through these hydrogels is
of utmost importance for cell viability and response, yet the translation
of diffusion coefficients into practical guidelines is not well established.
Here, we combined mathematical modeling, fluorescent recovery after
photobleaching, and hydrogel diffusion experiments on cell culture
inserts to provide a multiscale practical approach for diffusion.
We observed a dampening effect of the hydrogel that slowed the response
to concentration changes and the creation of a diffusion gradient
in the hydrogel by media refreshment. Our designed model combined
with measurements provides a practical point of reference for diffusion
coefficients in real-world culture conditions, enabling more informed
choices on hydrogel culture conditions. This model can be improved
in the future to simulate more complicated intrinsic hydrogel properties
and study the effects of secondary interactions on the diffusion of
analytes through the hydrogel
Optimization of Media Change Intervals through Hydrogels Using Mathematical Models
Three-dimensional cell culture in engineered hydrogels
is increasingly
used in tissue engineering and regenerative medicine. The transfer
of nutrients, gases, and waste materials through these hydrogels is
of utmost importance for cell viability and response, yet the translation
of diffusion coefficients into practical guidelines is not well established.
Here, we combined mathematical modeling, fluorescent recovery after
photobleaching, and hydrogel diffusion experiments on cell culture
inserts to provide a multiscale practical approach for diffusion.
We observed a dampening effect of the hydrogel that slowed the response
to concentration changes and the creation of a diffusion gradient
in the hydrogel by media refreshment. Our designed model combined
with measurements provides a practical point of reference for diffusion
coefficients in real-world culture conditions, enabling more informed
choices on hydrogel culture conditions. This model can be improved
in the future to simulate more complicated intrinsic hydrogel properties
and study the effects of secondary interactions on the diffusion of
analytes through the hydrogel
Future directions in managing aniridia-associated keratopathy
Congenital aniridia is a panocular disorder that is typically characterized by iris hypoplasia
and aniridia-associated keratopathy (AAK). AAK results in the progressive loss of corneal
transparency and thereby loss of vision. Currently, there is no approved therapy to delay or
prevent its progression, and clinical management is challenging because of phenotypic
variability and high risk of complications after interventions; however, new insights into
the molecular pathogenesis of AAK may help improve its management. Here, we review
the current understanding about the pathogenesis and management of AAK. We highlight
the biological mechanisms involved in AAK development with the aim to develop future
treatment options, including surgical, pharmacological, cell therapies, and gene therapies.This work was supported by the European Unionâs Cooperation on Science and Technology (COST) Program, under COST Action CA-18116 ANIRIDIA-NET: âAniridia: networking to address an unmet medical, scientific, and societal challenge; DFG German Research Council FOR 2240 (www.for2240.de) to C. C. and M. N.; the European Joint Programme on Rare Diseases (EJP RD 2020) to N. L., D. B. G., D. A. (AAK-INSIGHT); the Science Foundation Ireland Investigator Award (Grant 12/IA/1624), the European Regional Development Fund and the European Unionâs Horizon 2020 Research and Innovation Programme (Grant 814439) to T. R.; Academy of Finland (Grant 338988) to H. S.; the Dutch Research Council ZonMw TOP (Grant 91217058, VISION) to M. M. D., S. F. and V. L. S. L.peer-reviewe