Kanda Hideki, Ando Daigo, Hoshino Rintaro, Yamamoto Tetsuya, Suzuki Shogo, Shinohara Satoshi, Goto Motonobu
Department of Materials Process Engineering, Nagoya University, Nagoya 464-8603, Japan.
Department of Chemical Systems Engineering, Nagoya University, Nagoya 464-8603, Japan.
ACS Omega. 2021 May 14;6(20):13417-13425. doi: 10.1021/acsomega.1c01549. eCollection 2021 May 25.
Porcine aortic tissue was decellularized by subcritical dimethyl ether (DME) used as an alternative to the surfactant sodium dodecyl sulfate. The process included three steps. For the first step, lipids were extracted from the porcine aorta using subcritical DME at 23 °C with a DME pressure of 0.56 MPa. Next, DME was evaporated from the aorta under atmospheric pressure and temperature. The second step involved DNA fragmentation by DNase, which was primarily identical to the common method. For the third step, similar to the common method, DNA fragments were removed by washing with water and ethanol. After 3 days of DNase treatment, the amount of DNA remaining in the porcine aorta was 40 ng/dry-mg, which was lower than the standard value of 50 ng/mg-dry. Hematoxylin and eosin staining showed that most cell nuclei were removed from the aorta. These results demonstrate that subcritical DME eliminates the need to utilize surfactants.
猪主动脉组织通过使用亚临界二甲醚(DME)进行脱细胞处理,以此替代表面活性剂十二烷基硫酸钠。该过程包括三个步骤。第一步,在23°C、DME压力为0.56 MPa的条件下,使用亚临界DME从猪主动脉中提取脂质。接下来,在常压和常温下从主动脉中蒸发掉DME。第二步是通过脱氧核糖核酸酶(DNase)使DNA片段化,这与常规方法基本相同。第三步,与常规方法类似,通过用水和乙醇洗涤来去除DNA片段。经过3天的DNase处理后,猪主动脉中残留的DNA量为40 ng/干毫克,低于50 ng/毫克干重的标准值。苏木精和伊红染色显示,主动脉中的大多数细胞核已被去除。这些结果表明,亚临界DME无需使用表面活性剂。