Three copies of the size Order of a first 5 × 104 HBCEC were INCUBATEd with different concentrations of chemotherapeutic compounds, epothilone LY404039 A and B, epirubicin, doxorubicin, in a 96-well plate for 6 d at 37, 5% CO2. The determination of the ATP TCA was gem the manufacturer’s protocol with the untreated cells and cells treated with the Inhibitorl ATP solution with the standard maximum-ATP performed embroidered. After lysis of the tumor cells with an extraction buffer, the luminescence was luciferinluciferase in a fluorine / luminometer after addition of the reagent and the percentage of intact cells was measured using the software recovery. Results of ex vivo cultured tumor tissue of breast cancer patients after surgery was the growth of breast cancer cells as adh Brought pension human epithelial cells in combination and displayed a massive Verl EXTENSIONS cytoplasmic projections Similar to the normal morphology as described for human mammary epithelial cells.
Unlike HMEC growth in monolayer cultures showed HBCEC multilayer cell growth KU-55933 and were connected together by many desmosomes. Immunofluorescence showed a significant expression of cytokeratin green in all cultures HBCEC, epithelial demonstrates t satisfied as a contamination by other cell types such as fibroblasts. Zus USEFUL tests for prolyl-4-hydroxylase fibroblastspecific remained below the detection limit in cultures HBCEC. Co immunofluorescence analysis was carried out with the red mark vimentin, appears also in some T cells. Blue DAPI staining F Of the cores and one overlay image showing the expression of cytokeratin and vimentin collaboration in a variety of cells, which shows a different intracellular Re localization of intermediately Ren filaments.
The quantification of the expression of vimentin and cytokeratin cytometry showed that approximately 99% of the cytokeratin positive cells in which approximately 32% of these Bev lkerungsgruppe Both vimentin positive and cytokeratin-positive cells are showed. However, analysis by flow cytometry of desmin filaments, which are primarily in myoepithelial and myofibroblastic Zellph Phenotypes observed no detectable F Staining or culture. Although the amount of vimentin in different cultures may vary HBCEC k, Levels were detected by cytokeratin always at 95% or more. Moreover remained w While the expression of intermediaries Rfilamente was of prime Ren cultures of tumor cells to obtain stable long-term culture 34d shows one Rfilamente Much the same pattern of the intermediaries.
Taken together, these data almost exclusively Lich as epithelial cell population HBCEC. To cell surface Chenmarker during long-term culture of breast tumors evaluated, Bev HBCEC POPULATION analyzed after 176 days for CD24, CD44 and CD227, compared with a culture of the patient’s tumor days even after the 462nd Sun CD24 is expressed in 89% and 86% of the 176d HBCEC HBCEC 462D. In addition, the occurrence of CD44 was detectable in 94% of HBCEC 176d and 462D HBCEC 99%, suggesting little or no Ver Change both CD24 and CD44 in the tumor-term culture. In contrast, the expression of CD227 protein surface che Substantially from 52% to 88% HBCEC 176d 462D obtained in HBCEC Ht.