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Published on: June 26, 2018
[A prompt method to quantitative assay of alpha-Gal on pig cell surface without injury]
Ling-yu Zeng1, Sheng-tu Li, Jie Zhang
1Key Lab of Transplant Engineering and Transplant Immunology, West China Hospital, Sichuan university, Chengdu 610041, China.
Objective:
To develop a method for quantitative assay of alpha-Gal on pig cell surface.
Methods:
After centrifugation of the anticoagulated blood samples, the white blood cell layer was collected for addition of FITC-BSIB4 and analysis using FCM as well as Laser Scan Confocal Microscope (Bio-Rad).
Results:
With human blood group B as negative control, and mouse myeloma SP2/0 cells as positive control. the peripheral blood cells of two Chinese pigs were all positive, there were about 9.16 x 10(5) alpha-Galactosyl epitopes expressed on the cell-membrane of granulocytes 1.37 x 10(5) on the monocytes of Banna Minipig Inbred Line (BMI), and 1.16 x 10(6) on the granulocyte and 2.45 x 10(5) on the monocytes of Sichuan White Pig (SWP). The levels of alpha-Galactosyl expression on the surface of blood cells in BMI and SWP differed in various cells, the sequence from high to lower expression levels being on the platelets, granulocytes, monocytes, erthrocytes in each species of pig.
Conclusion:
The method is useful for quantitative assay of alpha-Gal on pig cell surface, and the assay may be used for alpha-Gal for on-site large-scaled screening of living pigs.
