کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8419901 | 1545762 | 2009 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
A novel method for isolation of human lung T cells from lung resection tissue reveals increased expression of GAPDH and CXCR6
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کلمات کلیدی
FBSGAPDHRT-PCRMicroarray - ریزآرایهLung - ریهfetal bovine serum - سرم جنین گاوFlow cytometry - فلوسیتومتریT lymphocytes - لنفوسیتهای TAlveolar macrophages - ماکروفاژهای آلوئولارreverse transcription polymerase chain reaction - واکنش زنجیره ای پلیمراز رونویسی معکوسPropidium iodide - پروتئین یدیدglyceraldehyde 3-phosphate dehydrogenase - گلیسرولیدید 3-فسفات دهیدروژناز
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
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چکیده انگلیسی
Lung T lymphocytes are important in pulmonary immunity and inflammation. It has been difficult to study these cells due to contamination with other cell types, mainly alveolar macrophages. We have developed a novel method for isolating lung T cells from lung resection tissue, using a combination of approaches. Firstly the lung tissue was finely chopped and filtered through a nylon mesh. Lymphocytic cells were enriched by Percoll density centrifugation and the T cells purified using human CD3 microbeads, resulting in 90.5% ± 1.9% (n = 11) pure lymphocytes. The T cell yield from the crude cell preparation was 10.8 ± 2.1% and viability, calculated using propidium iodide (PI) staining and trypan blue, was typically over 95%. The purification process did not affect expression of CD69 or CD103, nor was there a difference in the proportion of CD4 and CD8 cells between the starting population and the purified cells. Microarray analysis and real time RT-PCR revealed upregulation of GAPDH and CXCR6 of the lung T cells as compared to blood-derived T cells. This technique highly enriches lung T cells to allow detailed investigation of the biology of these cells.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 342, Issues 1â2, 15 March 2009, Pages 91-97
Journal: Journal of Immunological Methods - Volume 342, Issues 1â2, 15 March 2009, Pages 91-97
نویسندگان
C.E. Day, S.D. Zhang, J. Riley, T. Gant, A.J. Wardlaw, C. Guillen,