Essay Examples on Temple College

It was double diluted with RPMI tissue culture media

It was double diluted with RPMI tissue culture media layered on 3ml Ficoll hypaque Sigma USA and centrifuged at 1500rpm for 15 minutes The lymphocyte ring obtained was collected in a separate NUNC sterile tube containing 13 ml RPMI The contents of the tube were mixed well and centrifuged at 1000 rpm for 5min The supernatant was discarded and the pellet was dissolved in 1ml RPMIC RPMI containing 20 FCS The cell viability was checked using Trypan blue method Phytohaemagglutinin PHA was added at a concentration of 1µl per 1million cells 500 µl of the culture was added to each well of a 24 well NUNC plate The culture was incubated at 37 0 C in a 5 CO2 incubator for 24 hrs Addition of plant extract After 24hrs 1µl of IL 2 1 100 in RPMIC was added to each well along with 1µl of different dilutions 2 3 4 5 mg ml of the aqueous extract of C collinus One of the wells in which the standard drug AZT was added was used as the positive control The plates were incubated at 37 0 C in a 5 CO2 incubator for 72 hrs Different dilutions of the extracts were also added to wells with healthy lymphocytes and incubated at 37 0 C in a 5 CO2 incubator for 72 hrs Cytotoxicity assay was carried out using Trypan blue dye Mishell and Shiigi 1980 Measurement of p24 antigen expression


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