Biomedical subjects
Publications and source records attributed to B Bandemir.
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The cultivation of human keratinocytes is an appropriate experimental model for a variety of cell biological, pharmacological and biochemical investigations and the production of in vitro epidermis suitable for grafting since Rheinwald and Green (1975) have established this method. A procedure is described to isolate and culture human keratinocytes using different culture conditions. About 1-2 million epidermal cells could be prepared by trypsinization over night at 4 degrees C from 1-2 cm2 skin. Primary cell cultures seeded on mitomycin C treated NIH 3T3 fibroblasts reached subconfluence after 15-20 days when Eagle MEM was used as culture medium and after 10-15 days of culture using a mixture of Dulbecco's MEM and Ham F12 (3:1). In each case the media were supplemented with different growth factors. Subcultures were seeded on NIN 3T3 feeder cells as well as in collagen or fibronectin coated dish. Multilayered epidermal sheets developed with in 12-15 days of culture.
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The cumulative irritation power examined of some chemical disinfectants usual in the GDR. The recognition of irritation time (IT50) demonstrates in the animal experiment (pigs) and in probands with the epidemiological reality a good accordance.
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A simple in vitro screening method was established, which allows estimation of the effect of environmental products on cell alteration or irritative potential. Primary cell suspensions, prepared from the epidermis of guinea pig ears through trypsination (0.2%) over night at 4 degrees C, were used. The cell suspension (3 mill./ml) and test substances were mixed in equal portions (500 microliters) and incubated at room temperature for approximately 90 min. The degree of cell alteration was determined by the trypan blue exclusion test. Chemically defined substances as well as disinfectants were employed. The cell alteration was clearly dependent upon substance concentration as well as incubation time. On the basis of the mean alteration time (AT50) measured in vitro, and the mean irritation time (IT50) determined in the patch test in vivo, the substances were ranked according to strength of cell alteration or irritative potency in vitro and in vivo. A good correlation (Spearman's rank correlation, rs = 0.76) was found between the in vitro and in vivo results. Thus, this method is suited for the screening of chemical products especially in industrial laboratories.