Manoochehrabadi Tahereh, Sharifi Zohreh, Yari Fatemeh
Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
Med J Islam Repub Iran. 2019 Jun 13;33:55. doi: 10.34171/mjiri.33.55. eCollection 2019.
Membrane-derived microparticles (PMPs) are produced from platelets during activation, storage, and apoptosis. PMP can transfer some adhesion molecules such as CXCR4 to CXCR4-negative cells. In this study, the ability of PMPs to deliver CXCR4 molecule to CXCR4-null targets (Daudi, K562 and U937 cell line) was evaluated and the different concentrations of PMPs were examined to transfer CXCR4. In this experimental study, PMPs were prepared using serial centrifugations. After confirmation of PMP with flow cytometry, PMP concentration was evaluated using the Bradford method. CXCR4-negative cell lines (1×105 cells/ml) were cultured in RPMI1640 with 10% FBS and 1% antibiotic. PMPs in 7 different concentrations were added to cell culture plates and incubated for 1 hour at 37ºc and 5% CO2. The presence of CXCR4 on cells was analyzed by flowcytometry. In this study, characterization of PMPs and cell lines were done by flow cytometry. Then, the PMPs' ability to transfer CXCR4 to null cells (Daudi, K562 and U937 cell lines) was evaluated in 7 concentrations (10, 20, 50,125, 250, 500, 1000 μg/mL); incubation lasted for 1 hour. The best result of transferring CXCR4 by PMP was done in the concentration of 250µg/mL. PMPs in different concentrations can transfer CXCR4 to target cells. Also, the increase of PMPs concentration up to 250µg/mL can increase the CXCR4 presence on null cells.
膜衍生微粒(PMPs)是血小板在激活、储存和凋亡过程中产生的。PMP可以将一些黏附分子如CXCR4转移到CXCR4阴性细胞。在本研究中,评估了PMPs将CXCR4分子递送至CXCR4缺失靶细胞(Daudi、K562和U937细胞系)的能力,并检测了不同浓度的PMPs转移CXCR4的情况。在本实验研究中,通过连续离心制备PMPs。用流式细胞术确认PMP后,采用Bradford法评估PMP浓度。将CXCR4阴性细胞系(1×105个细胞/毫升)接种于含10%胎牛血清和1%抗生素的RPMI1640培养基中培养。将7种不同浓度的PMPs加入细胞培养板,于37℃、5%二氧化碳条件下孵育1小时。通过流式细胞术分析细胞上CXCR4的存在情况。在本研究中,通过流式细胞术对PMPs和细胞系进行了表征。然后,评估了7种浓度(10、20、50、125、250、500、1000μg/mL)的PMPs将CXCR4转移至缺失细胞(Daudi、K562和U937细胞系)的能力;孵育持续1小时。PMP转移CXCR4的最佳结果出现在250μg/mL浓度时。不同浓度的PMPs均可将CXCR4转移至靶细胞。此外,PMPs浓度增加至250μg/mL可增加缺失细胞上CXCR4的表达。