Hydrogel coated HIF-1 alpha overexpression lentiviral vector compound for repairing brachial plexus avulsion and preparation method
A technology of HIF-1 and lentiviral vector, which is applied in the fields of botany equipment and methods, biochemical equipment and methods, nervous system diseases, etc., can solve the problems such as undiscovered brachial plexus avulsion injury, and achieve the promotion of angiogenesis and movement Functional recovery, simple process method, effect of promoting nerve regeneration and functional recovery
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Embodiment 1
[0030] A method for preparing a hydrogel-wrapped HIF-1α overexpression lentiviral vector complex for repairing brachial plexus avulsion nerve injury, comprising the following steps:
[0031] Step 1, synthesis of HIF-1α overexpression lentiviral vector: pHBLV-CMVIE-IRES-Puro synthesized by Hanheng Biotechnology Co., Ltd. (Shanghai, China) was selected as the lentiviral vector for the overexpression of HIF-1α, wherein, HIF-1α primer sequences are as follows: ggatctatttttggggtgaattcgccaccATGGA GGGCGCCGGCG (forward primer); tagaactagtctcgaggaattcTC AGTTAACTTGATCCAAA (reverse primer);
[0032] Step 2, preparation of hydrogel stock solution: PF-127 sterile powder and water are configured into a suspension with a certain mass-to-volume ratio, and then the suspension is shaken until the hydrogel is fully dissolved to obtain a mixed solution, Then, after the mixed solution is filtered and sterilized through a filter membrane, a sterile PF-127 hydrogel stock solution is obtained;
[00...
Embodiment 2
[0035] A method for preparing a hydrogel-wrapped HIF-1α overexpression lentiviral vector complex for repairing brachial plexus avulsion nerve injury, comprising the following steps:
[0036] Step 1, synthesis of HIF-1α overexpression lentiviral vector: pHBLV-CMVIE-IRES-Puro synthesized by Hanheng Biotechnology Co., Ltd. (Shanghai, China) was selected as the lentiviral vector for the overexpression of HIF-1α, wherein, The HIF-1α primer sequence is as follows: ggatctatttttggggtgaattcgccaccATGGA GGGCGCCGGCG (forward primer); tagaactagtctcgaggaattcTC AGTTAACTTGATCCAAA (reverse primer); in this embodiment, the titer of the lentiviral vector is 2 × 10 8 TU / mL.
[0037] Step 2, preparation of hydrogel stock solution: PF-127 sterile powder and deionized water are configured into a suspension with a mass volume ratio of 20%, and then the suspension is placed on a horizontal shaker at 2°C to Shake at a speed of 65 rpm for 18 hours until the hydrogel is fully dissolved to obtain a mixed...
Embodiment 3
[0042] A method for preparing a hydrogel-wrapped HIF-1α overexpression lentiviral vector complex for repairing brachial plexus avulsion nerve injury, comprising the following steps:
[0043] Step 1, synthesis of HIF-1α overexpression lentiviral vector: pHBLV-CMVIE-IRES-Puro synthesized by Hanheng Biotechnology Co., Ltd. (Shanghai, China) was selected as the lentiviral vector for the overexpression of HIF-1α, wherein, The HIF-1α primer sequence is as follows: ggatctatttttggggtgaattcgccaccATGGA GGGCGCCGGCG (forward primer); tagaactagtctcgaggaattcTC AGTTAACTTGATCCAAA (reverse primer); in this embodiment, the titer of the lentiviral vector is 2 × 10 8 TU / mL.
[0044] Step 2, preparation of hydrogel stock solution: PF-127 sterile powder and deionized water are configured into a suspension with a mass volume ratio of 15%, and then the suspension is placed on a horizontal shaker at 0° C. Shake at a speed of 60 rpm for 24 hours until the hydrogel is fully dissolved to obtain a mixed ...
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