Is there any consensus with regard to so called recommended UVB dose used in in vitro experiments? From the literature review conducted by me so far it seems that different doses are used.
We would like to conduct an experiment to check the potential of our compounds against UVB induced skin ageing.
Hi Sabina. I work routinely in primary human skin cells (fibroblasts, keratinocytes and melanocytes). We use a bank of FS20 lamps that emit 75% UVB, 25% UVA and minimal UVC. To filter out the UVC, a Kodacel filter is placed between the lamps and any samples to be irradiated. In order to use an appropriate concentration of UVB, we use a UVB meter which gives readings. Taking an average of 10 readings allows me to determine the length of time I need to leave the samples in order to get the desired dose. Since not all lamps are exactly the same, it is difficult to know exactly what dose to use. Are you looking to do high-dose single irradiations or low-dose chronic treatments? For irradiating monolayer culture of cells, lose dose would be 25 or 50 mJ/cm2. Higher doses of UVB would be 75, 90, 105 or 135 mJ/cm2. Are you working with cell lines or actual tissue? Higher doses of UVB can cause the tissue samples to have poorer morphology when doing histological studies.
Hi Anne,
Thank you for your response. I thought that there should be a generally accepted protocol for UVB irradiation of cells. Well..how to compare the results when each researcher is using different doses?
I am using HaCat cell line.
We were thinking about using a dose which would allow us to study for example oxidative stress, cell senescence and not be very damaging.
Could anybody suggest some papers related to this issue?
Hi Sabina. I have attached a paper regarding UV and oxidative stress.