Project description:The purpose of this experiment was to obtain samples for mRNA analysis in IHH cells infected with human interferon alpha. Immortalized human hepatocyte (IHH) cells were seeded at 1 x 10^6 cells per well on 6 well plates 3 days before infection. Treatments were initiated by replacing culture medium (containing 10% FBS) with fresh medium containing 500 U/ml of human interferon α (PBL Biosciences). Mock-treated cells were given normal culture medium only. Mock and treated cells were incubated at 37°C until sample collection time points. Infected samples were collected in triplet; time-matched mocks were collected in triplet in parallel with infected samples. Time points: 6, 12, and 18 h post-infection
Project description:The purpose of this experiment was to obtain samples for microRNA analysis in IHH cells infected with human interferon alpha. Immortalized human hepatocyte (IHH) cells were seeded at 1 x 10^6 cells per well on 6 well plates 3 days before infection. Treatments were initiated by replacing culture medium (containing 10% FBS) with fresh medium containing 500 U/ml of human interferon α (PBL Biosciences). Mock-treated cells were given normal culture medium only. Mock and treated cells were incubated at 37°C until sample collection time points. Infected samples were collected in triplet; time-matched mocks were collected in triplet in parallel with infected samples. Time points: 6, 12, and 18 h post-infection
Project description:The transcriptional response to interferon alpha is cell type specific. To data, majority of investigations of interferon alpha induced gene expression have been made using immortalized or transformed cell lines underlining interferon's importance for treatment of cancer but omitting physiological relevance. Here in we have determined gene expression change in primary culture of hepatocytes after interferon alpha treatment. We used Affymetrix Rat Genome 230 2 microarrays to determine gene expression profiles in primary rat hepatocytes after interferon alpha treatment and untreated cells.
Project description:The transcriptional response to interferon alpha is cell type specific. To data, majority of investigations of interferon alpha induced gene expression have been made using immortalized or transformed cell lines underlining interferon's importance for treatment of cancer but omitting physiological relevance. Here in we have determined gene expression change in primary culture of hepatocytes after interferon alpha treatment. We used Affymetrix Rat Genome 230 2 microarrays to determine gene expression profiles in primary rat hepatocytes after interferon alpha treatment and untreated cells. Primary hepatocytes were chosen because they are most relevant to physiological state in intact liver. Hepatocytes was isolated from rat liver using collagenase treatment procedure and purified on percoll gradient. Next day after isolation primary hepatocytes culture was cultivated with (or without) 250u/ml rat interferon alpha for 3 and 6 hours. Experiment was performed in 3 biological replicates. cRNA preparation was performed according manufacturer's recommendation from 5ug of total RNA and gene expression profiles were determined.
