Early, middle, and late stages of neural cells from ovine embryo in primary cultures.

The utilization of neural cells in culture has importantly increased the knowledge of the nervous system biology. In most studies, the investigations are performed on biological materials coming from common laboratory animals and the extrapolation of the results to other animals is not easy. For some studies, such as developmental biology of the nervous system, prion disease investigations, or agronomical production, the utilization of ovine neural cell cultures presents many advantages. Unfortunately, there are few data on the conditions of culture of such cells. In the present work, we investigated simple ways to obtain neurons and astrocytes from sheep brain. Viable neuronal cell cultures were obtained from 40 to 50 day old fetuses. Their morphologies were quite similar to that of neurons from rodent or chick brain and they were labeled by antineurofilament antibodies. Stages older than 50 days of pregnancy were unable to give viable culture of neurons. The stages of 40 day old fetus to newborn lamb were able to give viable astrocyte cultures. The common protoplasmic astrocytes were obtained and they were labeled by antiglial fibrillary acidic protein antibodies. The astrocytes contained glycogen, thus looking like the common astrocytes from rodents. Neuronal or astroglial cultures can be derived from 26 day old embryos, but the cultures contained contaminating cells. Among the latter cells, there were undifferentiated cells which were flat and epitheloid and which were grouped as islets. These cells could be maintained in culture for a time duration over 7 months, even after two passages. They differentiated principally in astrocytes with a radial configuration. This work shows how some neural cells can be simply and easily cultured from sheep brain. For the first time, neurons were cultured from the sheep embryonic brain. Moreover, stem cells were cultured for more than 7 months and, finally, glycogen accumulation in sheep astrocytes was shown to be the same as that in rodent astrocytes. The oligodendrocyte culture was already documented. Thus, sheep can easily be used as well as other models for neural cell studies.