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A modification of tooth germ cultivation in vitro and in ovo
M Peterka1, V Mandys, R Peterková
1Institute of Experimental Medicine, Czechoslovak Academy of Sciences, Prague.
Abstract:
Mandibular molar anlages excised from 17-day mouse foetuses were cultured in vitro or in ovo (on the chorioallantoic membrane). In both cases, the explants were underlain either with a Millipore filter or with a piece of fibrin foam. Tooth germs were harvested after 7 days of cultivation and processed histologically. Spatial arrangement was highly preserved in the tooth germs cultured in vitro on fibrin foam. In vitro cultures on Millipore filters revealed significant flattening of tooth germs, caused especially by the collapse of enamel organ and the pulp. The cytodifferentiation of tooth germs cultured in vitro on both substrates (Millipore filter, fibrin foam) was characterized by the presence of odontoblasts, polarizing ameloblasts and predentine. The cytodifferentiation of tooth germs cultured in ovo on Millipore filters placed on chorioallantoic membrane was characterized by the presence of odontoblasts, ameloblasts, predentine, dentine and enamel. However, the flattening of these explants was identical with the changes of the explants cultured on Millipore filters in vitro. In ovo cultivation on the fibrin foam failed to bring satisfactory results.
Insights
Culturing mouse tooth germs on fibrin foam in vitro preserved their spatial arrangement. Other methods, particularly Millipore filters, caused significant flattening and incomplete development of tooth structures.
Area of Science:
- Developmental biology
- Oral biology
- Tissue engineering
Background:
- Tooth germ development is a complex process.
- In vitro and in ovo culture systems are used to study tooth development.
- Substrate choice can significantly impact tissue morphology and differentiation.
Purpose of the Study:
- To investigate the effect of different culture substrates (Millipore filter, fibrin foam) and environments (in vitro, in ovo) on the spatial arrangement and cytodifferentiation of mouse mandibular molar anlages.
- To evaluate the suitability of these culture systems for studying early tooth development.
Main Methods:
- Mandibular molar anlages from 17-day mouse foetuses were cultured.
- Cultures were performed in vitro or in ovo on the chorioallantoic membrane.
- Explants were supported by Millipore filters or fibrin foam.
- Histological processing was performed after 7 days of cultivation.
Main Results:
- In vitro culture on fibrin foam preserved the spatial arrangement of tooth germs.
- In vitro culture on Millipore filters resulted in significant flattening and collapse of the enamel organ and pulp.
- Cytodifferentiation, including odontoblasts, ameloblasts, and predentine, occurred in vitro on both substrates.
- In ovo culture on Millipore filters showed more advanced cytodifferentiation (dentine, enamel) but also exhibited flattening.
- In ovo culture on fibrin foam yielded unsatisfactory results.
Conclusions:
- Fibrin foam is a suitable substrate for maintaining the spatial integrity of cultured tooth germs in vitro.
- Millipore filters, in both in vitro and in ovo systems, lead to significant morphological distortion.
- While some cytodifferentiation occurs, the optimal conditions for both spatial arrangement and advanced differentiation require further investigation.