Moravvej Hamideh; Rad Mahnaz Mahmoudi; Toossi Parviz; Khorasani Mohammad Taghi; Mirzadeh Hamid
Volume 12, Issue 4 , 2009, , Pages 111-116
Abstract
Background: Fibroblasts are mesenchymal cells that can be readily cultured in the laboratory and play a significant role in epithelialmesenchymal interactions, secreting various growth factors and cytokines that have a direct effect on epidermal proliferation, differentiation and formation of extracellular ...
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Background: Fibroblasts are mesenchymal cells that can be readily cultured in the laboratory and play a significant role in epithelialmesenchymal interactions, secreting various growth factors and cytokines that have a direct effect on epidermal proliferation, differentiation and formation of extracellular matrix. They have been incorporated into various tissue-engineered and used for a variety of clinical applications, including the treatment of burns, chronic venous ulcers and several other clinical applications in dermatology and plastic surgery. Method: Isolated fibroblasts by the enzymatic process from foreskin were cultivated successively in a culture medium to establish cell banking. Foreskin and the last subcultured cells were checked for HBV, HCV, HIV, HSV I, HSV II, HTLV I, HTLV II, EBV, CMV, Treponema Pallidum, Mycoplasma sp. and Clamydia. The 1st, 5th and 10th subcultured cells were processed for immunocytochemistry studies using a panel of monoclonal antibodies including antibodies to MHC class I & II antigens for ensuring the elimination of superficial cell antigens during cultivation. Subcultured cells were karyotyped to find any chromosomal abnormalities. The best passages were chosen for culturing on silicone sheets provided by the Iran Polymer and Petrochemical Institute. Results: Evaluation for bacteria and viruses by molecular methods was negative. Karyotyping of cultured fibroblasts after the 10th passage showed some abnormalities. HLA expression was imperceptible in the cells obtained from the 10th sub-culture. The best passages were from 5th to 10th for banking and culturing on silicone sheets. Conclusion: Expression of HLA on fibroblast surfaces was diminished during subculturing. To prevent chromosomal abnormalities in fibroblast passaging, we should select the best colony that is expected to be chromosomally stable with the least antigenicity. In our study, the 5th to 10th sub-cultures were the best cells for the purpose of grafting and acceleration of the wound healing.
Rad Mahnaz Mahmoudi; Mohammadi Akram Mir Amin; Barton Richard C
Volume 11, Issue 1 , 2008, , Pages 17-20
Abstract
Background: Trichophyton rubrum (T. rubrum) is the most common cause of dermatophytosis of skin and nail tissue. Strain identification in Trichophyton rubrum is important for identification of strain-related differences in infectivity potential or transmissibility and epidemiological studies. PCR typing ...
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Background: Trichophyton rubrum (T. rubrum) is the most common cause of dermatophytosis of skin and nail tissue. Strain identification in Trichophyton rubrum is important for identification of strain-related differences in infectivity potential or transmissibility and epidemiological studies. PCR typing could determine whether the original isolate is responsible for re-infection or a new strain has been acquired.Methods: A minipreparation method for DNA from dermatophytes was used. Tandemly repetitive subelements (TRS-1 & TRS-2) of NTS region at ribosomal DNA of 23 T.rubrum isolates were amplified and the PCR products were separated by electrophoresis in 2% agarose gel (200 mA, 140 V), visualized by staining with ethidium bromide, and photographed.Results: On the basis of copy number of TRS-1 and TRS-2, 8 out of our 23 samples were type 2 & II, respectively. Six of them were type 3 & II, four isolates were type 1 & II, two isolates were type 4 & II, two isolates were type 1 & I and one isolate was type 5 & II.Conclusion: In this study, most of T. rubrum isolates were type 2 & II, dissimilar to European studies where type 1 & II has been the most common. The present study showed that 26.1% of Iranian isolates were type 1 in contrast with a previous study which has demonstrated a much lower prevalence in Asians (5%).