Dspe peg2000

Blank liposomes were firstly prepared according to the following procedures. Egg phosphatidylcholine (EPC) (Germany Lipoid company, Shanghai local agent, China), cholesterol and polyethylene glycol distearoylphosphosphatidylethanolamine (PEG₂₀₀₀-DSPE, NOF Corporation, Japan) (52:43:5, mmol/mmol) was dissolved by methanol in a pear-shaped flask. The methanol was evaporated to dryness under vacuum with a rotary evaporator (EYELA-1000S, EYELA Rikakikai Corporation, Tokyo, Japan), then hydrated with 250 mM ammonium sulfate by sonication in a water bath for 5 min. Suspensions were treated with an ultrasonic cell disruptor (JY92 cell sonicator, Ningbo Chibio-Technology Co., Ltd., Ningbo, China) for 3 min (200 W), and successively extruded through polycarbonate membranes (Millipore, Bedford, MA, USA) with pore size of 400 nm and 200 nm for 3 times, respectively. After extrusion, the suspensions were further dialyzed (12,000–14,000 molecular mass cutoff) against the phosphate buffered saline (PBS) (137 mM NaCl, 2.7 mM KCl, 8 mM Na₂HPO₄ and 2 mM KH₂PO₄, pH 7.4) for 3 times (each time 8 h) to obtain the blank liposomes.

We purchased DSPC, DSPG, and DSPE‐PEG2000 from NOF Corporation. Perfluoropropane (C₃F₈) was obtained from Takachiho Chemical Industrial Co. Ltd.
The cervical cancer HeLa cell line and endometrial cancer HEC1B cell line were purchased from ATCC. Cisplatin was purchased from Nichi‐Iko Pharmaceutical Company. Pegylated liposomal doxorubicin was purchased from Mochida Pharmaceutical Co., Ltd. Bevacizumab was obtained from Chugai Pharmaceutical Co., Ltd. Both DMEM and EMEM were purchased from Fujifilm Wako Pure Chemical Corporation. Fetal bovine serum was purchased from Thermo Fisher Scientific. Hematoxylin (Mayer’s hematoxylin; Wako) and 0.5% eosin Y ethanol solution were purchased from FUJIFILM Wako Pure Chemical Corporation. WST‐8 was purchased from Dojindo. Anti‐mouse CD31 Ab and anti‐mouse CD31 (PECAM‐1) were purchased from Dianova. Takara DAB substrate was purchased from Takara Bio Inc. BALB/c nude female mice and C57BL/6 mice were purchased from CLEA Japan, Inc. SP‐10 tubes were purchased from Natsume Seisakusho Co., LTD. Acuson Sequoia 512 (diagnostic US) was purchased from Siemens. An in situ apoptosis detection kit was purchased from Takara Bio Inc. All animal experiments were carried out in accordance with the Guidelines for the Care and Use of Laboratory Animals, as stated by The University of Tokyo.