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ePaper created 2017-11-24, 11:00:53 | version 1.38.0
| research photodamage of pulpa cells Photodamage of dental pulpa stem cells during 700 fs laser exposure Authors: Prof. Dr Karsten König & Dr Anton Kasenbacher, Germany Material and methods Cells Human stem cells from the dental pulp of adults (given by S. Gronthos, NIH, Bethesda, USA) were cultivated in -modifi ed Eagle’s Medium (MEM) by Gibco BRL Life Technologies (Paisley, Scotland) while adding 20 % FCS, 2 mM L-glutamine, 100 µM L-ascorbate-2-phosphate, 100 µ Penicillin and 100 µg/ml Streptomycin at 5 % CO2 and 37 °C in 25 ml and 75 ml cell culture fl asks (Greiner, Frickenhausen, Germany).1 The cells were treated for experiments and cultivation with 0.25 % trypsin, 5 mM glucose, 0.05% EDTA in PBS for 5 minutes at 37 °C. After hav- ing been thus detached from the base of the culture fl asks, the cells were incubated in cell chambers (MiniCeM, JenLab GmbH, Jena, Germany) for laser microscopy. Comparative studies were conducted on Chinese hamster ovary (CHO) cells, which are available in many international laboratories as reference cells. The CHO cells were incubated in Dulbeccos HAM-F12 Medium (Gibco BRL) with 10 % FCS, L-glutamine and an antibiotic mix of Penicillin, Streptomycin and Am- photericin B at 5 % CO2 and 37 °C. Trypsinisation cor- responded to that of the stem cells. Fig. 1: Femtosecond laser system with pulse-stretching unit. Laser-scanning microscope Sensor diode Objective 10 roots 4 2017 Mirrors with three- point support Beamer expander Auto- correlator Grating 2 on motorized translation stage Attenuator Fs laser Pulse strechter Grating 1 Mirror