TY - JOUR
T1 - Corneal epithelial–stromal fibroblast constructs to study cell–cell communication in vitro
AU - McKay, Tina B.
AU - Karamichos, Dimitrios
AU - Zieske, James D.
AU - Hutcheon, Audrey E.K.
AU - Guo, Xiaoqing
N1 - Funding Information:
This research was funded by the National Institutes of Health (NIH) and National EyeEyeInstitute (NEI),(NEI), R01EY005665, 5T32EY007145-20, and the NEI Core grant P30EY003790. In addition, tissues procured by NDRI with support from NIH 2U42 OD011158.
Funding Information:
Funding: This research was funded by the National Institutes of Health (NIH) and National Eye Institute (NEI), RR0011EEYY000055666655, ,5 5TT3322EEYY000077114455--2200,, aannddt thheeN NEEII CCoorree ggrraanntt PP3300EEYY000033779900.. IInn aaddddiittioionn, ,ttiissssuueess pprrooccuurreedd bbyyNNDDRRI I with support from NIH 2U42 OD011158.
Publisher Copyright:
© 2019 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2019/12
Y1 - 2019/12
N2 - Cell–cell communication plays a fundamental role in mediating corneal wound healing following injury or infection. Depending on the severity of the wound, regeneration of the cornea and the propensity for scar development are influenced by the acute resolution of the pro-fibrotic response mediated by closure of the wound via cellular and tissue contraction. Damage of the corneal epithelium, basement membrane, and anterior stroma following a superficial keratectomy is known to lead to significant provisional matrix deposition, including secretion of fibronectin and thrombospondin-1, as well as development of a corneal scar. In addition, corneal wounding has previously been shown to promote release of extracellular vesicles from the corneal epithelium, which, in addition to soluble factors, may play a role in promoting tissue regeneration. In this study, we report the development and characterization of a co-culture system of human corneal epithelial cells and corneal stromal fibroblasts cultured for 4 weeks to allow extracellular matrix deposition and tissue maturation. The secretion of provisional matrix components, as well as small and large extracellular vesicles, was apparent within the constructs, suggesting cell–cell communication between epithelial and stromal cell populations. Laminin-1β was highly expressed by the corneal epithelial layer with the presence of notable patches of basement membrane identified by transmission electron microscopy. Interestingly, we identified expression of collagen type III, fibronectin, and thrombospondin-1 along the epithelial–stromal interface similar to observations seen in vivo following a keratectomy, as well as expression of the myofibroblast marker, α-smooth muscle actin, within the stroma. Our results suggest that this corneal epithelial–stromal model may be useful in the study of the biochemical phenomena that occur during corneal wound healing.
AB - Cell–cell communication plays a fundamental role in mediating corneal wound healing following injury or infection. Depending on the severity of the wound, regeneration of the cornea and the propensity for scar development are influenced by the acute resolution of the pro-fibrotic response mediated by closure of the wound via cellular and tissue contraction. Damage of the corneal epithelium, basement membrane, and anterior stroma following a superficial keratectomy is known to lead to significant provisional matrix deposition, including secretion of fibronectin and thrombospondin-1, as well as development of a corneal scar. In addition, corneal wounding has previously been shown to promote release of extracellular vesicles from the corneal epithelium, which, in addition to soluble factors, may play a role in promoting tissue regeneration. In this study, we report the development and characterization of a co-culture system of human corneal epithelial cells and corneal stromal fibroblasts cultured for 4 weeks to allow extracellular matrix deposition and tissue maturation. The secretion of provisional matrix components, as well as small and large extracellular vesicles, was apparent within the constructs, suggesting cell–cell communication between epithelial and stromal cell populations. Laminin-1β was highly expressed by the corneal epithelial layer with the presence of notable patches of basement membrane identified by transmission electron microscopy. Interestingly, we identified expression of collagen type III, fibronectin, and thrombospondin-1 along the epithelial–stromal interface similar to observations seen in vivo following a keratectomy, as well as expression of the myofibroblast marker, α-smooth muscle actin, within the stroma. Our results suggest that this corneal epithelial–stromal model may be useful in the study of the biochemical phenomena that occur during corneal wound healing.
KW - Co-cultures
KW - Collagen
KW - Cornea
KW - Epithelium
KW - Extracellular vesicles
KW - Fibroblasts
KW - Stroma
UR - http://www.scopus.com/inward/record.url?scp=85078026887&partnerID=8YFLogxK
U2 - 10.3390/bioengineering6040110
DO - 10.3390/bioengineering6040110
M3 - Article
AN - SCOPUS:85078026887
SN - 2306-5354
VL - 6
JO - Bioengineering
JF - Bioengineering
IS - 4
M1 - 110
ER -