Shrimp lymphocytes and methods for their purification and methods for assessing innate immune responses in vitro
A technology of lymphocytes and purification methods, which is applied in the field of invertebrate lymphocytes and their purification, can solve the problems that shrimp cell lines have not been established, and achieve the effect of improving cell production, cell purity, and cell survival rate
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Embodiment 1
[0035] Example 1: Purification of Shrimp Lymphocytes
[0036] 1. Preparation Examples 1 to 6 and Preparation Comparative Examples 1 to 2
[0037] Preparation examples 1 to 6 and preparation comparative examples 1 to 2 are prepared anticoagulant agents according to Table 1, wherein preparation comparative examples 1 to 2 are anticoagulant agents of known formulations, and preparation examples 1 to 6 are several anticoagulant agents according to the present invention. Examples of anticoagulants. The above ingredients were dissolved in sterile water, and the pH was adjusted to 7.0 with 1N aqueous sodium hydroxide solution, and the bacteria were removed by filtration through a filter membrane with a pore size of 0.22 μm, and placed at 4°C for use.
[0038] Table 1
[0039]
preparation example 7
[0041] Preparation Example 7 and Preparation Comparative Example 3 prepared cell culture fluid according to Table 2, wherein Preparation Comparative Example 3 was a culture fluid with a known formula, and Preparation Example 7 was a cell culture fluid according to an embodiment of the present invention. The above ingredients were dissolved in sterile water, and the pH was adjusted to 7.0 with 1N aqueous sodium hydroxide solution, and the bacteria were removed by filtration through a filter membrane with a pore size of 0.22 μm, and placed at 4°C for use.
[0042] Table 2
[0043]
[0044] Table 2 shows the formulations of the cell culture fluids in Preparation Example 7 and Comparative Example 3. When preparing, add glucose to 800mL of sterilized water and dissolve completely, then add sodium chloride and 2 packs of Leibovitz’s L-15 powder [equivalent to 2 times the concentration (2X) of L-15]. After the above ingredients are completely dissolved, adjust the total volume to...
Embodiment 2
[0055] Example 2: Evaluating the Anti-Agglutination Effect of Anti-Agglutination Agents on Shrimp Lymphocytes
[0056] 1. Assess the anti-agglutination effect of anti-coagulant glucosamine or / and trypsin on shrimp lymphocytes
[0057] In this example, 18 white shrimps were used to evaluate the anti-coagulation and cell viability of shrimp lymphocytes with or without glucosamine or / and trypsin by using the anti-coagulant formulations of Preparation Examples 1 to 5 and Preparation Comparative Example 1 Impact.
[0058] Will be collected by point B (such as figure 1 Shown in Figure No. 103) obtained shrimp body fluid cells, quickly mixed with the anticoagulants of Preparation Examples 1 to 5 and Preparation Comparative Example 1 in the syringe, after mixing uniformly with a volume ratio of 1:1 to form a mixed solution, take 10 μL of the mixed solution, plus 90 μL of 0.5% trypan blue solution, calculate the number of cells and cell viability every 10 minutes, the results are as ...
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