Tissue reconstruction in 3D-spheroids from rodent retina in a motion-free, bioreactor-based microstructure.
While conventional rotation culture-based retinal spheroids are most useful to study basic processes of retinogenesis and tissue regeneration, they are less appropriate for an easy and inexpensive mass production of histotypic 3-dimensional tissue spheroids, which will be of utmost importance for future bioengineering, e.g. for replacement of animal experimentation. Here we compared conventionally reaggregated spheroids derived from dissociated retinal cells from neonatal gerbils (Meriones unguiculatus) with spheroids cultured on a novel microscaffold cell chip (called cf-chip) in a motion-free bioreactor. Reaggregation and developmental processes leading to tissue formation, e.g. proliferation, apoptosis and differentiation were observed during the first 10 days in vitro (div). Remarkably, in each cf-chip micro-chamber, only one spheroid developed. In both culture systems, sphere sizes and proliferation rates were almost identical. However, apoptosis was only comparably high up to 5 div, but then became negligible in the cf-chip, while it up-rose again in the conventional culture. In both systems, immunohistochemical characterisation revealed the presence of Müller glia cells, of ganglion, amacrine, bipolar and horizontal cells at a highly comparable arrangement. In both systems, photoreceptors were detected only in spheroids from P3 retinae. Benefits of the chip-based 3D cell culture were a reliable sphere production at enhanced viability, the feasibility of single sphere observation during cultivation time, a high reproducibility and easy control of culture conditions. Further development of this approach should allow high-throughput systems not only for retinal but also other types of histotypic spheroids, to become suitable for environmental monitoring and biomedical diagnostics.
Rieke
Matthias
M
Institute of Zoology, Darmstadt University of Technology, Schnittspahnstrasse 13, D-64287, Darmstadt, Germany.
Gottwald
Eric
E
Weibezahn
Karl-Friedrich
KF
Layer
Paul Gottlob
PG
D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis; D019149:Bioreactors; D049109:Cell Proliferation; D002478:Cells, Cultured; D005849:Gerbillinae; D048091:Guided Tissue Regeneration; D007150:Immunohistochemistry; D008904:Miniaturization; D009038:Motion; D012160:Retina / Q000166:cytology*; D018874:Spheroids, Cellular