Department of Diabetes, Endocrinology and Metabolism, Beckman Research Institute, City of Hope, Duarte, CA.
Department of Surgical Intensive Care Unit, Beijing Chao-Yang Hospital, Capital Medical University, Beijing, People's Republic of China.
Transplantation. 2020 Jun;104(6):e144-e150. doi: 10.1097/TP.0000000000003191.
Current histological methods cannot accurately determine the survival rate of human pancreatic islets following portal vein infusion. This is due, in part, to the low number of infused islets relative to the whole liver. In this study, we assessed the ability of confocal laser scanning microscopy (CLSM) to track human islets posttransplantation.
Immunodeficient mice were transplanted with human islets. Following engraftment, animals were euthanized, livers procured, and human islet β cells immunofluorescently labeled with an insulin-specific antibody and evaluated by CLSM. A calibration curve comparing the area of insulin + hepatic islet β cells to the number of human islets collected was developed. Levels of human C-peptide were measured in transplant recipients to determine islet function.
The short-term survival rate of islet transplants was defined as y = 0.0422x + 2.7008, in which x is human islet number and y is liver islet β cell area. Employing CLSM, human islets were detected in immunofluorescent labeled murine liver tissue sections posttransplantation. The β cell-relative area of human islets in 500 islet equivalent (IEQ) specimens was 20.21 ± 1.16 mm and in 1000 IEQ specimens 39.4 ± 2.23 mm posttransplantation. Human islet posttransplant survival rates were 82.9 ± 5.50% (500 IEQ group) and 86.9 ± 5.28% (1000 IEQ group).
These data indicate that CLSM can be employed to quantify and characterize pancreatic human islets after transplantation to murine livers.
目前的组织学方法无法准确确定门静脉输注后人类胰岛的存活率。这部分是由于输注的胰岛数量相对于整个肝脏而言较少。在这项研究中，我们评估了共聚焦激光扫描显微镜 (CLSM) 在移植后追踪人类胰岛的能力。
免疫缺陷小鼠接受人胰岛移植。在移植物植入后，处死动物，获取肝脏，并使用胰岛素特异性抗体对人胰岛 β 细胞进行免疫荧光标记，然后通过 CLSM 进行评估。建立了一个比较胰岛素+肝胰岛 β 细胞面积与人胰岛数量的校准曲线。测量移植受者中的人 C 肽水平以确定胰岛功能。
胰岛移植的短期存活率定义为 y = 0.0422x + 2.7008，其中 x 是人胰岛的数量，y 是肝胰岛 β 细胞的面积。通过 CLSM，在移植后可以在免疫荧光标记的鼠肝组织切片中检测到人胰岛。在 500 个胰岛当量 (IEQ) 标本中，人胰岛的 β 细胞相对面积为 20.21 ± 1.16 mm，在 1000 IEQ 标本中为 39.4 ± 2.23 mm。人胰岛移植后的存活率分别为 82.9 ± 5.50%（500 IEQ 组）和 86.9 ± 5.28%（1000 IEQ 组）。
这些数据表明，CLSM 可用于定量和表征移植到鼠肝后的胰腺人胰岛。
