Composition for treating canine parvovirus disease and preparation method thereof
A technology of canine parvovirus and its composition, which is applied in the field of composition for treating canine parvovirus disease and its preparation, can solve the problems that there are no specific drugs for parvovirus, and achieve the effects of promoting production, improving quality of life, and inhibiting parvovirus
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Embodiment 1
[0028] Example 1 - Preparation of canine adipose-derived mesenchymal stem cells
[0029] (1) Isolation and culture of canine adipose-derived mesenchymal stem cells
[0030] Take a dog (healthy adult dog, no rabies virus, canine distemper virus, canine parainfluenza virus, canine infectious hepatitis virus, canine parvovirus, canine coronavirus, fungal and bacterial infection.) about 1cm of abdominal fat tissue 3 , washed with PBS containing 5% v / v double antibody (containing a mixture of penicillin (10,000IU) and streptomycin (10,000μg / mL) at a working concentration of 100 times.), then cut into pieces, and transferred to a centrifuge Add collagenase type I at a concentration of 1 mg / mL to the tube, digest in a 37°C incubator for 1 hour, filter the digested solution through a 100-mesh cell sieve, centrifuge the filtrate for 5 minutes, discard the supernatant, and add canine adipose mesenchyme Stem cell culture solution (DMEM containing 10% v / v fetal bovine serum and 1% wt L-g...
Embodiment 2
[0042] Example 2- Preparation of culture supernatant of canine adipose-derived mesenchymal stem cells
[0043] When the canine adipose-derived mesenchymal stem cells isolated and cultured according to Example 1 were cultured to a confluence of about 80%, washed with PBS for 3 times, added serum-free medium (basic medium DMEM, Gibco), placed at 37 ° C, 5 After culturing in a constant temperature incubator with % carbon dioxide for 72 hours, the serum-free cell supernatant was collected into a centrifuge tube, and centrifuged at 3000 g at 4° C. for 30 min to remove cell residues in the cell supernatant. Sterile filtration was performed through a 0.22 μm filter membrane after centrifugation. Obtain a pure cell supernatant, which is the culture supernatant of canine adipose-derived mesenchymal stem cells.
[0044] After testing, the protein concentration of the canine adipose-derived mesenchymal stem cell culture supernatant is 1-8 mg / mL.
[0045]The canine adipose-derived mesen...
Embodiment 3
[0046] Example 3 - Preparation of a composition for the treatment of canine parvovirus disease
[0047] The composition for treating canine parvovirus disease of the present invention comprises canine adipose-derived mesenchymal stem cells prepared according to Example 1, canine adipose-derived mesenchymal stem cell culture supernatant prepared according to Example 2 and canine interferon. The canine adipose-derived mesenchymal stem cell culture supernatant and canine interferon are mixed evenly according to the amount, and then the canine adipose-derived mesenchymal stem cells are added to obtain the composition for treating canine parvovirus disease of the present invention.
[0048] In the composition, the volume ratio of canine adipose mesenchymal stem cell culture supernatant to canine interferon is 1:2 to 5:1.
[0049] In this composition, according to the sum of the volumes of canine adipose-derived mesenchymal stem cell culture supernatant and canine interferon, 10±5×1...
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