Application of doxycycline ad medicine for promoting cell proliferation
A cell proliferation, doxycycline technology, applied in the direction of drug combination, tetracycline active ingredients, etc., can solve problems such as inability to relieve pulmonary hypertension
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Embodiment 1
[0045] Example 1. Proliferation-promoting effect of doxycycline on mouse thymus medullary epithelial cell line MTEC1
[0046] When doxycycline is used as the inducer of the Tet-on (tetracycline-induced promoter system) system to induce the expression of related genes in the MTEC1 cell line, use 3 Cell proliferation was measured by the H-TdR incorporation assay. The specific steps are as follows: After washing the cells of the mouse thymus medullary epithelial cell line MTEC1 in the logarithmic growth phase, they were suspended in 5×10 cells with DMEM complete medium. 4 / ml, inoculated in a 96-well culture plate, 100 μl per well, respectively added doxycycline 0.1, 1, 10 and 100 μg / ml to the cell culture, and set up a control group (without doxycycline). Each treatment group made 6 duplicate wells, and each concentration treatment group was cultured for 24, 48, and 72 hours respectively, and 0.5 μCi / well was added 12 hours before cell harvest. 3 H-TdR, continue to culture to ...
Embodiment 2
[0053] Example 2. Proliferation-promoting effect of doxycycline on other cells
[0054] Contrary to our results, many literatures reported that doxycycline can inhibit the proliferation and induce apoptosis of many tumor cells, activated T and B cells. Under the same experimental conditions as in Example 1, we used 3 The effect of doxycycline on cell lines from different sources was detected by H-TdR incorporation method. Such as image 3 and Figure 4 As shown, doxycycline can promote the proliferation of HEK293, CTX-3, TEC1C8, TNC-R3.1 and mouse fresh thymic stromal cells, but the effect on these cells is weaker than that of MTEC1; on the contrary, it can Inhibits Con A-activated T cells, T lymphoma cell lines (Jurkat, C2 and EL-4) and LPS-activated B cells; as image 3 As shown, doxycycline has no obvious pro-proliferative effect on HUVEC cell lines. In addition, these effects are dose-dependent. (Note: ConA is concanavalin A, LPS is lipopolysaccharide)
Embodiment 3
[0055] Example 3. Morphological observation of the effect of doxycycline on promoting the proliferation of MTEC1 cells
[0056] In order to further confirm the role of doxycycline in promoting the proliferation of MTEC1 cells, we used the method of morphological observation. 3×10 5 / ml (about 3.12×10 4 / cm 2 ) MTEC1 cells were inoculated in 6-well cell culture plates, 1ml per well, added different concentrations of doxycycline and cultured for 72 hours, and a blank control was set up at the same time, with three replicate wells in each group, observed and photographed under an inverted microscope. Such as Figure 5 As shown, after MTEC1 cells were cultured with different concentrations of doxycycline for 7 days, the cells in the control group shown in group A died; in group B, a small number of cells cultured with 0.1 μg / ml doxycycline began to die; in group C, 1 μg / ml doxycycline The cells cultured with 1 ml doxycycline continued to grow; in group D, the cells cultured wi...
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