首页> 美国卫生研究院文献>International Journal of Molecular Sciences >Modification of MCF-10A Cells with Pioglitazone and Serum-Rich Growth Medium Increases Soluble Factors in the Conditioned Medium Likely Reducing BT-474 Cell Growth
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Modification of MCF-10A Cells with Pioglitazone and Serum-Rich Growth Medium Increases Soluble Factors in the Conditioned Medium Likely Reducing BT-474 Cell Growth

机译:用吡格列酮和富含血清的生长培养基修饰MCF-10A细胞会增加条件培养基中的可溶性因子从而可能降低BT-474细胞的生长

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摘要

In the present study, we aimed to preincubate MCF-10A cells with pioglitazone and/or serum-rich growth media and to determine adhesive and non-adhesive interactions of the preincubated MCF-10A cells with BT-474 cells. For this purpose, the MCF-10A cells were preincubated with pioglitazone and/or serum-rich growth media, at appropriate concentrations, for 1 week. The MCF-10A cells preincubated with pioglitazone and/or serum-rich growth media were then co-cultured adhesively and non-adhesively with BT-474 cells for another week. Co-culture of BT-474 cells with the preincubated MCF-10A cells, both adhesively and non-adhesively, reduced the growth of the cancer cells. The inhibitory effect of the preincubated MCF-10A cells against the growth of BT-474 cells was likely produced by increasing levels of soluble factors secreted by the preincubated MCF-10A cells into the conditioned medium, as immunoassayed by ELISA. However, only an elevated level of a soluble factor distinguished the conditioned medium collected from the MCF-10A cells preincubated with pioglitazone and serum-rich growth medium than that with pioglitazone alone. This finding was further confirmed by the induction of the soluble factor transcript expression in the preincubated MCF-10A cells, as determined using real-time PCR, for the above phenomenon. Furthermore, modification of the MCF-10A cells through preincubation did not change the morphology of the cells, indicating that the preincubated cells may potentially be injected into mammary fat pads to reduce cancer growth in patients or to be used for others cell-mediated therapy.
机译:在本研究中,我们旨在用吡格列酮和/或富含血清的生长培养基预孵育MCF-10A细胞,并确定预孵育的MCF-10A细胞与BT-474细胞的粘附和非粘附相互作用。为此,将MCF-10A细胞与吡格列酮和/或富含血清的生长培养基(适当浓度)预孵育1周。然后将与吡格列酮和/或富含血清的生长培养基预温育的MCF-10A细胞与BT-474细胞粘附和非粘附共培养另外一周。 BT-474细胞与预孵育的MCF-10A细胞的粘附性和非粘附性共培养减少了癌细胞的生长。通过ELISA免疫测定,预培养的MCF-10A细胞对BT-474细胞生长的抑制作用可能是通过增加预培养的MCF-10A细胞分泌到条件培养基中的可溶性因子水平来产生的。然而,与单独使用吡格列酮相比,只有升高水平的可溶性因子才能区分从预先与吡格列酮和富含血清的生长培养基一起孵育的MCF-10A细胞中收集的条件培养基。对于上述现象,如通过实时PCR确定的,通过在预孵育的MCF-10A细胞中诱导可溶性因子转录物表达进一步证实了这一发现。此外,通过预孵育对MCF-10A细胞的修饰不会改变细胞的形态,这表明预孵育的细胞可能会被注入乳腺脂肪垫中,以减少患者的癌症生长或用于其他细胞介导的治疗。

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