Search PubMed⌕ Search

Biomedical subjects

J Farrant

Publications and source records attributed to J Farrant.

At least 91 records · Page 5Linked to original sources
PubMed1977

Cryopreservation of mouse peritoneal macrophages.

A method for the preservation of mouse peritoneal macrophages is described. Using 5% dimethylsulphoxide with either a cooling rate of 1 degree C/min or two-step cooling with 10 min interruption at -30 degrres C allows 80% survival as judged by pinocytic activity. An important finding was that the cells must be handled at 0 degrees C both before and after freezing.

Animals↗
PubMed1977

Recovery of infective Schistosoma mansoni schistosomula from liquid nitrogen: a step towards storage of a live schistosomiasis vaccine.

Successful recovery of infective schistosomula of Schistosoma mansoni following storage at -196 degrees C is reported. The technique involves a two-step cooling procedure--slow cooling (0.65 degrees C min-1) to an intermediate temperature of -28 degrees C, followed by rapid cooling into liquid nitrogen (10,000 degrees C min-1). Rewarming (10,000 degrees C min-1) and rapid dilution to remove the cryoprotectant (17.5% methanol) yielded motile organisms some of which developed to adult worms in mice after intramuscular injection. The percentage of schistosomula developing to adult worms was small (0.44%), but is a significant step towards storage of trematode larvae and of a live attenuated vaccine for schistosomiasis.

Animals↗
PubMed1977

Structural and functional aspects of biological freezing techniques.

The cooling procedures used to prepare samples for ultrastructural examination at low temperatures often differ markedly from those used to recover optimal function of cells on thawing. The implications of these differences are reviewed. Damage and alteration to the structure and function of the cells may be caused by the high concentrations of cryoprotective agents such as glycerol or dimethyl sulphoxide (DMSO) often added to reduce ice crystal artefacts. Under the rapid cooling conditions commonly employed for structural studies, these additives are not cryoprotective; low rates of cooling are necessary for them to be effective. Rapidly cooled cells that contain intracellular ice are only injured during rewarming so their structure may be as yet unaltered by any damaging effects at low temperatures. Most cells able to recover on thawing are grossly shrunken at low temperatures but since they are potentially functional they are of interest structurally. These cryobiological principles are illustrated with freeze-fracture, freeze substitution and functional assays. The cell types chosen were Chlorella sp. and mammalian tissue culture cells.

Animals↗
PubMed1977

Survival of chlamydiae after cooling to -196 degrees C.

Factors influencing the survival of chlamydiae after freezing were reexamined. From the data presented, it is suggested that preservation of laboratory-grown chlamydiae is best achieved through the use of sucrose as the cryoprotective agent, in the presence of 10% serum. Dimethyl sulfoxide and glycerol are more toxic. The period of exposure to sucrose before freezing must be kept as short as possible and be at 4 degrees C rather than at room temperature. The rate of cooling during freezing in sucrose is not important; however, cooling at a rate slower than 1 degrees C/min should be avoided. Since chlamydial survival is increased by rapid thawing, the volume of the sample should be kept to a minimum. Thawed suspensions should be inoculated onto cell monolayers without delay. The application of these methods may increase the proportion of stored clinical specimens in which chlamydiae can be found.

Blood↗
PubMed1977

Preservation of intraerythrocytic forms of malarial parasites by one-step and two-step cooling procedures.

Ring, trophozoite, and schizont stages of Plasmodium knowlesi were cooled in dimethyl sulfoxide either by direct immersion in liquid nitrogen or by a two-step method in which the cells were held at temperatures slightly below 0 degrees C for different lengths of time before they were cooled to -196 degrees C. After the direct plunge treatment, thawed trophozoites and schizonts were found to be extensively damaged. Their survival was markedly increased by holding them at -31 degrees C for 30 min before plunging them into liquid nitrogen. Freeze-substitution showed that cells cooled by the two-step procedure were grossly shrunken and had relatively few intracellular ice cavities. Large amounts of ice formed in trophozoites and schizonts preserved by direct immersion in liquid nitrogen. The two-step protocols investigated did not improve the survival of ring-stage parasites, 25-50% of which survived rapid cooling to -196 degrees C. Infected cell agglutination tests were carried out with frozen and thawed schizonts. Variant specificity was demonstrated with cells that had been plunged directly to -196 degrees C, but cells cooled by the two-step method tended to agglutinate spontaneously.

Animals↗