Meng Fu-Tao, Huang Mei, Fan Fang-Fang, Shao Feng, Wang Chao, Huang Qiang
Department of General Surgery, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei, Anhui Province, People's Republic of China,
Anhui Province Key Laboratory of Hepatopancreatobiliary Surgery, Anhui Provincial Hospital, Hefei, Anhui Province, People's Republic of China,
Cancer Manag Res. 2019 Feb 14;11:1533-1539. doi: 10.2147/CMAR.S192354. eCollection 2019.
This study explored a simple, high-yield method for isolating quiescent human pancreatic stellate cells (PSCs) to provide sufficient and reliable raw materials for PSC-related studies.
Single-cell suspensions were prepared from normal human pancreatic tissue specimens using the gentleMACS tissue processor, which enhanced the yield and viability of the suspensions. Percoll density gradient centrifugation was then performed to isolate quiescent normal PSCs (NPSCs). Cell viability was determined by trypan blue staining, and the states of the NPSCs were determined by autofluorescence and oil red O staining. The purity of human activated PSCs (APSCs) was determined by immunofluorescence assays.
The yield of NPSCs was ~(2.75±0.65)×10 cells/g. The maximum cell viability was 92%, whereas the maximum cell purity was 95%.
The method employed in this study to isolate PSCs is a simple, high-yield and stable method that is worth popularizing.
本研究探索了一种简单、高产的方法来分离静止的人胰腺星状细胞（PSC），为PSC相关研究提供充足且可靠的原材料。
使用gentleMACS组织处理器从正常人胰腺组织标本中制备单细胞悬液，这提高了悬液的产量和活力。然后进行Percoll密度梯度离心以分离静止的正常PSC（NPSC）。通过台盼蓝染色测定细胞活力，通过自发荧光和油红O染色确定NPSC的状态。通过免疫荧光测定法确定人活化PSC（APSC）的纯度。
NPSC的产量约为（2.75±0.65）×10⁵个细胞/克。最大细胞活力为92%，而最大细胞纯度为95%。
本研究中用于分离PSC的方法是一种简单、高产且稳定的方法，值得推广。
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