Comparison of two methods used to culture and purify rat retinal Mller cells

• Published in: 国际眼科杂志：英文版 2013 (6), p.778-784
• Main Author: Wei-Tao Song Xue-Yong Zhang Si-Qi Xiong Dan Wen Jian Jiang Xiao-Bo Xia
• Format: Article
• Language: English
• Subjects: cell; culture; Mller; passage; primary; purification; retinal; trypsinization
• ISSN: 2222-3959; 2227-4898

AIM:To study two methods for culturing and purifying Sprague-Dawley(SD)rat retinal Muller cells and determine which one is better.METHODS:The passage culture method of Muller cells was respectively carried out by complete pancreatic enzyme digestion method and repeated incomplete pancreatic enzyme digestion method.After culturing retinal cells for one month through these two methods,fluorescence-activated cell sorter(FACS),RT-PCR,and immunohistochemistry technology were performed to examine the enrichment and purity of Muller glial cells,and carried out two-sample approximate t test using SSPS 13.0 to further compare the Muller cell positive rate in both methods.RESULTS:The statistical results showed that the purity of Muller cells was 83.2%±5.16%in group A,and the purity was 98.5%±1.08%in group B.The two-sample approximate t test analysis demonstrated that the difference between group A and group B was statistically significant(t=-9.178,P<0.005).The results clearly exhibited a difference between the purity of Muller cells cultured by the complete pancreatic enzyme digestion method(group A)and the repeated incomplete pancreatic enzyme digestion method(group B).CONCLUSION:Compared with the complete pancreatic enzyme digestion method,this novel method was more efficient and a higher purity of Muller cells could be obtained using this approach.