کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8420074 | 1545772 | 2008 | 11 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Unbiased analysis, enrichment and purification of thymic stromal cells
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کلمات کلیدی
TECTSCthymic stromal cellsingle positive - تک مثبتThymus - تیموسDouble negative - دو برابر منفی استdouble positive - دو مثبتepithelial cells - سلول بافت پوششی یا اپیتلیومDendritic cell - سلول دندریتیکstromal cells - سلول های استروماthymic epithelial cell - سلول های اپیتلیال تومیکFlow cytometry - فلوسیتومتریPurification - پاکسازی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
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چکیده انگلیسی
The microenvironment of the thymus consists of functionally discrete niches composed of distinct stromal cell subsets. Clinically relevant changes affecting T-cell differentiation occur within these niches with age and injury caused by irradiation and chemotherapy treatments. The study of thymic stromal cells has been hampered by the technical difficulty in isolating significant numbers of this important population. Here we present an improved protocol for enzymatic isolation of stromal cells that enables comparative flow cytometric analyses and their purification for downstream cellular or molecular analysis. Fractions analyzed throughout enzymatic digestion of the thymus revealed that various stromal subsets are isolated at characteristic intervals. This highlights the importance of pooling all cells isolated from the thymus for numerical and phenotypic analysis to avoid biased representation of subpopulations. We also describe refined magnetic bead separation techniques that yield almost pure preparations of CD45â stroma. Sorting of these suspensions using defined markers enabled purification of the major epithelial subsets, confirmed by keratin staining and PCR analysis. This three-step procedure represents a rapid, reproducible method for the unbiased purification of the stromal cells that direct thymic T-cell differentiation.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 329, Issues 1â2, 1 January 2008, Pages 56-66
Journal: Journal of Immunological Methods - Volume 329, Issues 1â2, 1 January 2008, Pages 56-66
نویسندگان
Daniel H.D. Gray, Anne L. Fletcher, Maree Hammett, Natalie Seach, Tomoo Ueno, Lauren F. Young, Jade Barbuto, Richard L. Boyd, Ann P. Chidgey,