Retention of bone cell viability in mouse calvarial explants after cryopreservation

Newborn mouse calvaria, cyropreserved at -196 degrees C in serum-free medium containing dimethyl-sulfoxide, were compared to unpreserved explants for bone cell viability by [3H]thymidine uptake. Other explants were studied using autoradiography to compare the histological appearance of the cryopreserved and control unpreserved explant sites of cellular localization of [3H]thymidine. After short-term cryopreservation, calvarial bone cells, including less differentiated osteoprogenitor cells, survived as indicated by their incorporation of the DNA precursor. With culture continuing for up to 24 hr after thawing and in the continuous presence of [3H]thymidine, additional labeled thymidine was incorporated, indicating that the proliferative ability of explant cells persists after cryopreservation. Cryopreserved bone explants did not, however, incorporate the same amount of labeled thymidine as did controls at each time point studied. These events, as measured quantitatively and observed by autoradiography of the tissue, indicate that newborn calvarial bone cell proliferation in vitro continues after cryopreservation. The large surface:mass ratio of the tissue and its proportionate volume of calcified matrix apparently permits it to behave as an isolated cell population with regard to the diffusion of the cryoprotectant and thermal conductivity, thus permitting the retention of explant viability.     

DIFFERENTIAL INCORPORATION OF [3H]LYSINE INTO VISUAL CORTEX PROTEIN FRACTIONS DURING FIRST EXPOSURE TO LIGHT

—A resolution of the enhancement of protein synthesis in the visual cortex of rats during first exposure to light (Richardson and Rose , 1972) was achieved by polyacrylamide gel electrophoresis using a double-labelling technique. Differential incorporation of lysine was established between exposed and control animals in two fractions of the soluble proteins and seven fractions of the insoluble proteins. This suggests that exposure to a new experience of this type involves a specific effect on protein synthesis, rather than a general stimulation across all fractions.