I am currently working with RPMI8866 but some of my experiments have hit a block owing to the non-adherent nature of these cells. Thus I am looking into any adherent alternatives.
Saumya Nigam!
I think, that you should continue with RPMI and begin to use
the JY cells.
Most mononuclear leukocytes and cell lines express the integrin a4D1 (VLA-4) heterodimer. In this study, we have used Northern blotting and immunoprecipitation experiments to demonstrate that a B lymphoblastoid cell line (JY) expressed the integrin f17 subunit in association with a4. These a4B7-positive JY cells bound poorly or not at all to VLA-4 ligands (soluble form of vascular cell adhesion molecule 1 (sVCAM-1) and the CS1 region of fibronectin). In contrast, a positive variant of JY cells (selected to express a mixture of a4b1 and a“&) bound avidly to VLA-4 ligands, and this binding was completely inhibitable by anti-a4 and anti-dl monoclonal antibodies. Thus, expression appears to be a critically important component of VLA4-mediated binding to its ligands. After either JY or JY-& cells were stimulated for 15 min with the phorbol ester 12-O-tetradecanoylphorbol-13-acetate, the majority of adhesion to VCAM or fibronectin remained a4- and bI-dependent, but a low amount of adhesion to sVCAM- 1 or fibronectin became a4-dependent, B1-independent, thus suggesting a role for a4&. In summary, we have found (i) that a4@7 makes little or no contribution to fibronectin or VCAM-1 binding on unstimulated JY cells, (ii) that a“@7 perhaps makes a minor contribution to ligand binding on 12-0-tetradecanoylphorbol-13-acetate-stimulated cells, and (iii) that a4B1 is the functionally dominant VCAM-1 and fibronectin receptor even when expressed in relatively low amounts compared to a‘&.
The JY cell line is an Epstein–Barr virus (EBV)-immortalised B cell lymphoblastoid line. Cells express HLA class-I A2 and class-II DR. It is a suspension cell line, although cells are known to grow in clumps. The growth medium is RPMI 1640, 10% fetal calf serum and 1% L-glutamine.
I hope, that these data are usefull for you. If not, please ask again, and I will help you as much as possible.
Sincerely yours!
Dr. Bratko Filipič
Email: [email protected]
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