Cryopreservation, hepatocyte monolayers and DMSO shock

Cryopreservation of primary hepatocyte monolayers may provide a means of long-term storage of the cells for in vitro studies of xenobiotic metabolism and toxicity. This talk focuses on cryopreservation, hepatocyte monolayers and DMSO shock

Cryopreservation of rat hepatocyte monolayers: cell viability and cytochrome P450 content in post-thaw cultures

Cryopreservation of primary hepatocyte monolayers may provide a means of long-term storage of the cells for in vitro studies of xenobiotic metabolism and toxicity. Rat hepatocytes can be stored at −70 °C as simple monolayers attached to collagen-coated dishes, and post-thaw cultures can be continued for up to 72 h. Throughout this post-thaw period viability of the cells was demonstrated by retention of intracellular fluorescence after exposure to carboxyfluorescein diacetate (CFDA) and examination by confocal laser scanning microscopy (CLSM). CLSM images revealed an uneven distribution of CFDA-derived fluorescence within hepatocytes post-thaw, particularly in Williams' E medium, indicating generation and retention of carboxyfluorescein within the intracellular organelles. The membranes of the intracellular organelles appear to be less sensitive to freeze/thaw damage than the cell membrane. Viability was not compromised with storage for up to 28 days at −70 °C. Cytochrome P450 content was retained in post-thaw culture to a similar extent as in non-frozen cultures. Cryopreserved rat hepatocyte monolayers may provide a useful in vitro model for studying xenobiotic metabolism and toxicity

Growth and viability of osteoblasts on hydroxyapatite and tri-calcium phosphate ceramics: the effect of collagen coating

Summary of paper describing how Hydroxyapatite (HA) and tricalcium phosphate (TCP) ceramics form the basis for bioartificial bone replacement materials. They lack mechanical strength and cannot at present be considered as replacements for loadbearing areas. We have investigated the characteristics of two ceramic materials, HA and a 60:40 composite of HA:TCP, which are being developed for facial bone reconstruction. The growth and viability of an immortalized cell line, FFC, derived from neonatal rat calvaria, was investigated when cultured on the materials for 7 days