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Yuki Maekawa, Akishi Onishi, Naoshi Koide, Kiyoshi Suzuma, Michiko Mandai, Takashi Kitaoka, Masayo Takahashi; Extracellular matrices preferable to neurite outgrowth from retinal organoids differentiated from mouse embryonic stem cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):1769.
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© ARVO (1962-2015); The Authors (2016-present)
As an additional tool to investigate neurite outgrowth of RGC, we recently reported a method to induce neurite outgrowth from retinal organoid differentiated from mouse and human embryonic stem cells. In the previous study, we observed that biological coating with matrigel served important roles for neuritogenesis. However, because matrigel consist of plural components of extracellular matrix, a key element for neuritogenesis from retinal organoid has remained to be explored.
Retinal organoids were differentiated from Fstl4::Venus mouse embryonic stem cells as Eiraku et al. had described previously. Fstl4 expression has been reported in some specific subtype of RGC in embryonic mouse retina. After isolating optic vesicles from retinal organoids, vesicles were settled into adhesion culture on containers coated with matrigel or single components of ECM; 100% matrigel, 10% matrigel, collagen type 1, collagen type 4, laminin, and fibronectin. The rate of vesicle adhesion and the incidence of processus were evaluated at day 1 and 3 after placement respectively.
The rates of vesicle adhesion on collagen type 1-coat was significantly lower than those on the others (adjusted p value<0.01%, Fisher’s exact test, adjusting the false discovery rate using the Benjamini-Hochberg procedure). The incidences of processus on collagen type 1, collagen type 4 and fibronectin induced processus at the significantly lower rate compared 10% and 100% matrigel and laminin (adjusted p value<0.01% for each). Neurites indicated different morphology on laminin-coated wells.
It was suggested that laminin has a crucial effect for adhesion and neurite outgrowth from retinal organoid derived from mouse embryonic stem cells. The morphological difference may be caused by another components in matrigel and requires further investigation.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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