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2 cm) was obtained aseptically to isolate stem cells. The biopsy was cut into thin pieces and treated with trypsin at 4° C overnight (cold trypsin method) to obtain a single-cell suspension. The cells were seeded at a density of 3×104 cells/cm2 onto a preformed mitomycine-C treated 3T3 cell as feeder layer in DMEM medium supplemented with 10% fetal bovine serum (FBS) and other special supplements. Clonogenic keratinocytes divided and colonies quickly expanded and pushed away the 3T3 feeder layer cells, which then detached from the culture vessel and eliminated with medium changes. Primary cultures were usually subcultured when the cells were in exponential growth phase .Colonies of keratinocytes were expanded and after 7-10 days fused and formed a coherent stratified epithelium. Confluent cultured epithelia were detached enzymatically as coherent sheets from the surface of the culture flasks and transferred onto petrolatum- impregnated gauze.Histological studies of cultured epithelium were also carried out.
Results: In our experience from 1 cm2 of skin sample, 2,5- 4×106 cells were obtained. It resulted in keratinocytes suspensions which consisted at least 90% single cells. Cultured keratinocytes proliferated and after 8-10 days became confluent. The area of cultured epithelium detached from T-25 and T-75 culture flasks was approximately 12-15 cm2 and 35-40 cm2 respectively. Histological studies showed that 10-day old cultured epithelium had 3-4 cell layers consisting of small basal cells and big scquamous cells with large nucleus. Also in the basal layer few melanocytes with melanin pigments in the cells cytoplasm were found.The 20-day old cultured epithelium had 8-10 layers consisting of small and round basal cells, scquamous cells and 2-3 layers of keratinized cells.
Conclusion: Culture of keratinocyte stem cells could result in multilayer epithelium that creates a good cosmetic appearance upon transplantation. This could re-generate an epidermis that is resistant to trauma and infections. It can be considered as an appropriate substitution in skin loss conditions.
Keywords: Epidermal Sheets, Skin Adult Stem cells, Keratinocyte.
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37/8 C°), signs of allergy, traumatic LP (bloody), and unusual local bleeding.
Results
In this study, 118 males and 84 females underwent 623 general anesthetic procedures with a median of 3 procedures per patient. Intraoperative period complications occurred in 48 of total 623 procedures (7.7 %). The most common complications were traumatic LP, bradycardia and decrease in arterial oxygen saturation which occurred in 25, 6 and 6 cases, respectively. Postoperative period complications occurred in 74 (11.9%) cases. The most common complications were vomiting, agitation and headache, decrease O2 saturation and bradycardia.
Conclusion
General anesthesia by propofol and fentanyl may be a good choice for short-term painful procedures in children undergoing treatment for bone marrow aspiration/biopsy and intratechal injection.
Key Words: Embryonic stem cells; Bone Marrow, Biopsy.]]>
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5%).
Conclusion:
In result, moreover theses agent affected on different organs, they destroyed cells. It can be suggested the loss of these cells may have a role in lake of regeneration of damaged tissues.
Keyword: Adipose tissue, Crack, Crystal, Mesenchymal stem cells.
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