Cellular proliferation assays

A p-anilyl-substituted an.va-titanocene 48 and titanocene 3 were tested on the growth of a wide variety of tumour cells in vitro on a panel 36 human tumour cell lines containing 14 different tumour types investigated in a cellular proliferations assay [34]. Titanocene 3 showed a significantly higher cytotoxic activity than the ansa-compound and reached, on average over the whole cell panel, the activity of cisplatin within a factor of 4. Nevertheless, there were three main targets for titanocene 3 identified, which are pleura mesothelioma, uterine and renal-cell... 

Figure 5.2 Sequences of important regions of Abl kinase, highlighting sites of resistance mutations identified from clinical tumor samples. Several mutations including Y253F-H, E255 K-V, and T315I have a > 10-fold impact on imatinib potency when tested in a Ba-F3 cellular proliferation assay.31 Only T315I retains such a significant resistance effect on the second generation inhibitors dasatinib and nilotinib.

The cellular proliferation assay is used to measure the effects of various washing solutions on cell viability see Note 5). 

Different formulations of saponin-adjuvanted vaccines were tested for humoral and cell-mediated responses in mice. Many antigens, often including hen egg albumin (ovalbumin, OVA) as the antigen to be tested, have been used for immunizations, enzyme immunoassays, antigen-specific CTL and cellular proliferation assays [31]. 

Triciribine, also known as Akt/protein kinase B signaling inhibitor-2 (API-2) compound 52, Fig. 9), was identified by screening the National Cancer Institute Diversity Set chemical library (1992 compounds). The screen was performed using a cell-based proliferation assay with PKB/1 transformed NIH3T3 cells, and triciribine scored positive in this assay with an IC50 value of 50 nM [ 196]. Although the compound blocks the cellular phosphorylation... 

Chemicals and pharmaceuticals may be tested for their capacity to induce skin sensitization [49]. The potency of a xenobiotic or pharmaceutical compound to induce delayed type hypersensitivity (DTH) or contact dermatitis (CD) may be tested in the so called local lymph node assay (LENA) in which the induction of an immune response in lymph nodes is determined after local (skin) exposure [49-51]. The induction of cellular proliferation in draining lymph nodes is measured by determining the... 

In the neutral red (cell viability) and total protein (cell proliferation) assays, cells are treated with various concentrations of a test substance in petri or multiwell dishes after a period of exposure, the substance is washed out of the medium. (An analytical reagent is added in the case of protein measurements.) Neutral red is a supravital dye, which accumulates in the lysosomes of viable, uninjured cells, and it can be washed out of cells, which have been damaged. In the protein test, Kenacid blue is added and reacts with cellular protein. Controlled cells are dark blue killed cells are lighter colored. The IC50 (the concentration which inhibits by 50%) is determined the test can be rapidly performed with automation. However, materials must be solubilized into the aqueous cell media for analysis. For many test materials this will require large dilutions which eliminate properties of the materials which cause irritation. 

Characteristic of immune function evaluations, there is considerable diversity in the approaches to proliferation assays, from the experimental design to the analytical method. Experimental procedures necessitate sterile technique and cell culture expertise to ensure accurate assessment of the cellular response to a particular stimulant. Culture conditions vary depending on the cell source and type of stimulant, but generally are conducted at 5% CO2,37 °C for 48 to 96 hours. The source of lymphocytes may be from peripheral blood, spleen. 

Interleukin-2 Human recombinant lL-2 aldesleukin, proleukin des-alanyl-1, serine-125 human lL-2) differs from native lL-2 in that it is not glycosylated, has no amino terminal Ala, and has an Ser substituted for the Cys at amino acid 125. The potency of the preparation is represented in International Units in a lymphocyte proliferation assay such that 1.1 mg of recombinant lL-2 protein equals 18 million International Units. Aldesleukin has the following in vitro biologic activities of native lL-2 enhancement of lymphocyte proliferation and growth of lL-2-dependent cell lines enhancement of lymphocyte-mediated cytotoxicity and killer cell activity and induction of interferon-7 activity. In vivo administration of aldesleukin in animals produces multiple immunologic effects in a dose-dependent manner. Cellular immunity is profoundly activated with lymphocytosis, eosinophilia, thrombocytopenia, and release of multiple cytokines e.g., TNF-a, lL-1, interferon-7). Aldesleukin is indicated for the treatment of adults with metastatic renal cell carcinoma and melanoma. Administration of aldesleukin has been associated with serious cardiovascular toxicity resulting from capillary leak syndrome, which involves loss of vascular tone and leak of plasma proteins and fluid into the extravascular space. Hypotension, reduced organ perfusion, and death may occur. An increased risk of disseminated infection due to impaired neutrophil function also has been associated with aldesleukin treatment. 

Proliferation assays on matched sensitive and resistant cell hnes Transport assays in monolayers of cells expressing P-gp A colorimetric assay for P-gp s ATPase activity Cellular assays using fluorescent or pro-fluorescent P-gp substrates... 

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