A marker for detection or auxiliary detection of retinal cone cells and application thereof
A technology of cone cells and retina, applied in the field of biomedicine, can solve problems such as the inability to apply animal models or cell model identification
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Embodiment 1
[0083] Example 1. Discovery and application of retinal cone cell markers
[0084] (1) Acquisition of cynomolgus monkey single-cell sequencing libraries (including cynomolgus monkey retinal sequencing libraries)
[0085] Schematic diagram of the construction and analysis of cynomolgus monkey single-cell sequencing library, see figure 1 .
[0086] 1. Obtaining Cynomolgus Monkey Cell Suspension
[0087] According to the average life expectancy of cynomolgus monkeys and its corresponding relationship with human age, the young group consists of 8 cynomolgus monkeys aged 4-6 (half male and female), and 8 cynomolgus monkeys (half male and female) aged 18-21 The monkeys made up the old group. All cynomolgus monkeys underwent physical examination, including the measurement of basic parameters (such as body mass index (BMI)), the detection of biochemical indicators (such as blood routine, urine routine), and the examination of important internal organs such as brain MRI and heart. U...
Embodiment 2
[0133] Example 2, the application of protein RBP4 in detecting whether the sample to be tested contains retinal cone cells
[0134] 1. Immunofluorescence detection
[0135] 1. Prepare slices of samples to be tested
[0136] If the sample to be tested is the retina, it is prepared according to the method of (2)-2 of Example 1.
[0137] If the sample to be tested is retinal cells, it is prepared according to the conventional method of cell sheet preparation.
[0138] The samples to be tested were retinal tissues from 3 young cynomolgus monkey individuals and 3 old cynomolgus monkey individuals.
[0139] 2. Frozen sections (approximately 8 μm) were placed on positively charged glass slides and stored at -80°C.
[0140] 3. Take the slices, first wash them with PBS buffer, then place them in pH 6.0, 10 mM sodium citrate buffer, and heat the slices at 98°C for 3 times (5 min each time) in microwave, for the purpose of antigen retrieval.
[0141] 4. After completing step 3, after...
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