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Int J Ophthalmol. 2013 Dec 18;6(6):778-84. doi: 10.3980/j.issn.2222-3959.2013.06.07. eCollection 2013.

Comparison of two methods used to culture and purify rat retinal Müller cells.

Author information

1
Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha 410008, Hunan Province, China.

Abstract

AIM:

To study two methods for culturing and purifying Sprague-Dawley (SD) rat retinal Müller cells and determine which one is better.

METHODS:

The passage culture method of Müller 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 Müller glial cells, and carried out two-sample approximate t test using SSPS 13.0 to further compare the Müller cell positive rate in both methods.

RESULTS:

The statistical results showed that the purity of Müller 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 Müller 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 Müller cells could be obtained using this approach.

KEYWORDS:

passage; primary culture; purification; retinal Müller cell; trypsinization

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