Clothier R H, Bourne N
School of Biomedical Sciences, Faculty of Medicine and Health Sciences, University of Nottingham, Nottingham, NG7 2UH UK.
J Biol Phys. 2003 Jun;29(2-3):179-85. doi: 10.1023/A:1024492725782.
Primary human keratinocytes can be driven,in vitro, to differentiate, viaactivation of transglutaminases, by raisingthe culture medium calcium concentrationabove 1 mM. This results intransglutaminase regulated cross linking ofspecific amino acids with resultantcornified envelope formation. Thedifferentiation was monitored via theincorporation of fluorescein cadaverineinto the cornified envelops. Thisdifferentiation assay was combined withassessment of reductive capacity ofresazurin, as a measure of cellactivity/viability.One primary aim is to assess the effects ofTHz radiation on human skin, since medicalimaging of the body through the skin isenvisaged.Human keratinocytes, at passage 2 fromisolation, were grown to confluence, andtransported in a buffered salt solution at22 (°)C. The exposure to the THz sourcewas for 10, 20 or 30 minutes at roomtemperature.No donor specific inhibition or stimulationof cell activity, compared with non-exposedcells, was noted following exposure in therange 1 to 3 THz, at up to 0.45J/cm(2).The differentiation also occurred in anormal way, for exposed and non-exposedcells, with the FC incorporation increasingbetween day 3 and day 8, as previouslynoted.
原代人角质形成细胞在体外可通过将培养基钙浓度提高到1 mM以上,经转谷氨酰胺酶激活而被驱动分化。这会导致转谷氨酰胺酶调节特定氨基酸的交联,从而形成角质包膜。通过将荧光素尸胺掺入角质包膜来监测分化情况。该分化测定与刃天青还原能力的评估相结合,作为细胞活性/活力的一种衡量指标。一个主要目的是评估太赫兹辐射对人体皮肤的影响,因为设想通过皮肤对身体进行医学成像。从分离开始传代2次的人角质形成细胞生长至汇合,然后在22℃的缓冲盐溶液中运输。在室温下,对太赫兹源的暴露时间为10、20或30分钟。在1至3太赫兹范围内、高达0.45J/cm(2)的暴露后,与未暴露细胞相比,未观察到供体特异性的细胞活性抑制或刺激。如前所述,暴露和未暴露细胞的分化也以正常方式发生,荧光素尸胺掺入量在第3天至第8天增加。