Open Access Highly Accessed Research article

Regulation of in vitro human T cell development through interleukin-7 deprivation and anti-CD3 stimulation

Ekta S Patel1, Starlyn Okada1, Kevin Hachey1, Li-jun Yang2, Scott K Durum3, Jan S Moreb4 and Lung-Ji Chang15*

Author Affiliations

1 Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Gainesville, FL, 32610, USA

2 Department of Pathology, Immunology and Laboratory Medicine, College of Medicine, University of Florida, Gainesville, FL, 32610, USA

3 Section of Cytokines and Immunity, National Cancer Institute, Frederick, MD, 32610, USA

4 Department of Medicine, College of Medicine, University of Florida, Gainesville, FL, 32610, USA

5 Department of Molecular Genetics and Microbiology, University of Florida, 1600 SW Archer Road, ARB R1-252, Gainesville, FL, 32610, USA

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BMC Immunology 2012, 13:46  doi:10.1186/1471-2172-13-46

Published: 16 August 2012

Additional files

Additional file 1:

Optimized PCR condition for Rag2 and TREC genomic DNA amplification.A, Titration curves of cloned Rag2 and TREC templates of known concentrations. The standardization condition was applied to create a log dilution series for each assay. The coefficient of correlation for Rag2 were R = 0.9993, and for TREC R = 0.9992, indicating equal amplification of the templates over a range of input DNA concentrations. B, The equal and comparable slopes established for Rag2 and TREC PCR.

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Additional file 2:

Surface phenotype analysis of thein vitrodifferentiated CD4 T cells. The T cells developed from adult BM HPCs in the LmDL1-FL7 co-culture following IL-7 withdrawal and anti-CD3/CD28 stimulation were analyzed for various surface markers as shown by flow cytometry, in comparison with control PBMCs.

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