Preparation method and application of targeting chondrocyte drug delivery system
A chondrocyte and delivery system technology, applied in the fields of genetic engineering and biomedicine, can solve the problem of low therapeutic potential of cell-targeted delivery of drug molecules, and achieve the effects of controlling the destruction of articular cartilage, promoting cartilage repair, and slowing down degradation.
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Embodiment 1
[0025] The present invention provides a technical solution: a preparation method and application of a chondrocyte-targeted drug delivery system, the method comprising the following steps:
[0026] S1: Construct a plasmid, and use the chondrocyte-carrying affinity peptide (CAP) to be fused and expressed on the exosome membrane protein (lamp2b) as a targeted genetically engineered exosome that can target chondrocytes;
[0027] S2: Screen stable cell lines, express plasmid CAP-lamp2b in DC cells, and screen stable cell lines;
[0028] S3: Extract cartilage-affinity-targeted exosomes, use exosome-free medium to expand cells stably expressing CAP-lamp2b, collect cell supernatant, and perform differential ultracentrifugation to obtain exosomes. 1. Design of lamp2b Primer, using mouse cDNA as a template, PCR amplification obtains the lamp2b fragment, PCR (polymerase chain reaction) purification;
[0029] 2. Cloning the lamp2b gene after step 1 PCR purification and recovery into the ...
Embodiment 2
[0040] The present invention provides a technical solution: a preparation method and application of a chondrocyte-targeted drug delivery system, the method comprising the following steps:
[0041] S1: Construct a plasmid, and use the chondrocyte-carrying affinity peptide (CAP) to be fused and expressed on the exosome membrane protein (lamp2b) as a targeted genetically engineered exosome that can target chondrocytes;
[0042] S2: Screen stable cell lines, express plasmid CAP-lamp2b in DC cells, and screen stable cell lines;
[0043] S3: Extract cartilage-affinity-targeted exosomes, use exosome-free medium to expand cells stably expressing CAP-lamp2b, collect cell supernatant, and perform differential ultracentrifugation to obtain exosomes. 1. Design of lamp2b Primer, using mouse cDNA as a template, PCR amplification obtains the lamp2b fragment, PCR (polymerase chain reaction) purification;
[0044] 2. Cloning the lamp2b gene after step 1 PCR purification and recovery into the ...
Embodiment 3
[0055] The present invention provides a technical solution: a preparation method and application of a chondrocyte-targeted drug delivery system, the method comprising the following steps:
[0056] S1: Construct a plasmid, and use the chondrocyte-carrying affinity peptide (CAP) to be fused and expressed on the exosome membrane protein (lamp2b) as a targeted genetically engineered exosome that can target chondrocytes;
[0057] S2: Screen stable cell lines, express plasmid CAP-lamp2b in DC cells, and screen stable cell lines;
[0058] S3: Extract cartilage-affinity-targeted exosomes, use exosome-free medium to expand cells stably expressing CAP-lamp2b, collect cell supernatant, and perform differential ultracentrifugation to obtain exosomes. 1. Design of lamp2b Primer, using mouse cDNA as a template, PCR amplification obtains the lamp2b fragment, PCR (polymerase chain reaction) purification;
[0059] 2. Cloning the lamp2b gene after step 1 PCR purification and recovery into the ...
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