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Cell Journal، جلد ۱۳، شماره Supplement، صفحات ۰-۰
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عنوان انگلیسی P-61: Isolation of Non-Adherent Multipotent Human Dermal Fibroblast
چکیده انگلیسی مقاله Objective: Dermal fibroblast as a major resident of connective tissue has attracted more attention in the past few years and application of these highly fast growing cells in the several fields has been intensively studied. Contrary to common belief, recent studies have revealed fibroblasts are not fully differentiated cells and there are small populations of multipotent dermal fibroblasts which can differentiate into mesodermal, ectodermal and endodermal progenies in vitro. The main purpose of this investigation was to obtain single-cell-derived clone from dermal fibroblasts. Materials and Methods: Dermal fibroblasts were isolated from fresh human foreskin of donors aged from 1 to 3 months. The samples were washed and subcutaneous tissues were removed carefully and after adding 0.5% Dispase II at 37°C for 2 hours the epidermal layers were removed and dermal parts were digested with 0.1% Collagenase at 37°C. The released cells were collected every 20 minutes and passed through a 70μm cell strainer, centrifuged and resuspend in complete growth medium (high-glucose DMEM+ 10% FBS+ 1% penicillin/streptomycin. The cells were then seeded in 96-well tissue culture plates by limiting dilution. Results: After 7 days some small colonies appeared in wells where contain small number of seeded cells and they have become sizeable in 2 weeks and also budding structures have been seen on surfaces of them. Conclusion: Although culture conditions of current study have been prepared based on previous reports to isolate multipotent dermal fibroblasts which are adherent cells, we observed several colonies in suspension. Given that subcutaneous and epidermal layer had been removed earlier, presumably these clones belong to dermis but further investigation should be done to characterize them accurately
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/1593
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