PAMAM-cRGD mediating efficient siRNA delivery to spermatogonial stem cells
Abstract Background Spermatogonial stem cells (SSCs) are the cornerstone of sperm production and thus perpetual male fertility. In clinics, transplantation of patient’s own SSCs into testes is a promising technique to restore fertility when male germ cells have been depleted by gonadotoxic therapies...
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| Format: | Article |
| Language: | English |
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BMC
2019-12-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-019-1506-4 |
| _version_ | 1818380718494973952 |
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| author | Tianjiao Li Qiwen Chen Yi Zheng Pengfei Zhang Xiaoxu Chen Junna Lu Yinghua Lv Shiguo Sun Wenxian Zeng |
| author_facet | Tianjiao Li Qiwen Chen Yi Zheng Pengfei Zhang Xiaoxu Chen Junna Lu Yinghua Lv Shiguo Sun Wenxian Zeng |
| author_sort | Tianjiao Li |
| collection | DOAJ |
| description | Abstract Background Spermatogonial stem cells (SSCs) are the cornerstone of sperm production and thus perpetual male fertility. In clinics, transplantation of patient’s own SSCs into testes is a promising technique to restore fertility when male germ cells have been depleted by gonadotoxic therapies. Auto-transplantation of genetically modified SSCs even has the potential to treat male infertility caused by genetic mutations. However, SSCs are refractory to transfection approaches. Poly(amidoamine) (PAMAM) dendrimers have the unique three-dimensional architecture, surface charge, and high density of surface groups that are suitable for ligand attachment, thereby facilitating target delivery. The goal of this study was to elucidate whether PAMAM dendrimers can efficiently deliver short interfering RNAs (siRNAs) to SSCs. Methods and results We introduced cyclic arginine-glycine-aspartic acid (cRGD) peptides to the fifth generation of PAMAM dendrimers (G5) to generate PAMAM-cRGD dendrimers (G5-cRGD). The characterization of G5-cRGD was detected by Fourier transform infrared spectroscope (FTIR), transmission electron microscope (TEM), and the Cell Counting Kit-8 (CCK-8) assay. Confocal microscopy and flow cytometry were used to evaluate the delivery efficiency of siRNA by G5-cRGD to SSCs. The results showed that G5-cRGD encompassing siRNA could self-assemble into spherical structures with nanoscale size and possess high transfection efficiency, excellent endosomal escape ability, and low cytotoxicity, superior to a commercial transfection reagent Lipofectamine® 2000. Moreover, we demonstrated that G5-cRGD efficiently delivered siRNAs and triggered gene silencing. Conclusions This study thus provides a promising nanovector for siRNA delivery in SSCs, facilitating the future clinical application of SSC auto-transplantation with genetically modified cells with a hope to cure male infertility that is caused by genetic disorders. |
| first_indexed | 2024-12-14T02:23:08Z |
| format | Article |
| id | doaj.art-01cfbe9379874ccda33b011128eee447 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-14T02:23:08Z |
| publishDate | 2019-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-01cfbe9379874ccda33b011128eee4472022-12-21T23:20:27ZengBMCStem Cell Research & Therapy1757-65122019-12-0110111510.1186/s13287-019-1506-4PAMAM-cRGD mediating efficient siRNA delivery to spermatogonial stem cellsTianjiao Li0Qiwen Chen1Yi Zheng2Pengfei Zhang3Xiaoxu Chen4Junna Lu5Yinghua Lv6Shiguo Sun7Wenxian Zeng8Key Laboratory for Animal Genetics, Breeding and Reproduction of Shaanxi Province, Key Laboratory for Animal Biotechnology, Ministry of Agriculture of China, College of Animal Science and Technology, Northwest A&F UniversityShaanxi Key Laboratory of Natural Products & Chemical Biology, College of Chemistry & Pharmacy, Northwest A&F UniversityKey Laboratory for Animal Genetics, Breeding and Reproduction of Shaanxi Province, Key Laboratory for Animal Biotechnology, Ministry of Agriculture of China, College of Animal Science and Technology, Northwest A&F UniversityKey Laboratory for Animal Genetics, Breeding and Reproduction of Shaanxi Province, Key Laboratory for Animal Biotechnology, Ministry of Agriculture of China, College of Animal Science and Technology, Northwest A&F UniversityKey Laboratory for Animal Genetics, Breeding and Reproduction of Shaanxi Province, Key Laboratory for Animal Biotechnology, Ministry of Agriculture of China, College of Animal Science and Technology, Northwest A&F UniversityShaanxi Key Laboratory of Natural Products & Chemical Biology, College of Chemistry & Pharmacy, Northwest A&F UniversityShaanxi Key Laboratory of Natural Products & Chemical Biology, College of Chemistry & Pharmacy, Northwest A&F UniversityShaanxi Key Laboratory of Natural Products & Chemical Biology, College of Chemistry & Pharmacy, Northwest A&F UniversityKey Laboratory for Animal Genetics, Breeding and Reproduction of Shaanxi Province, Key Laboratory for Animal Biotechnology, Ministry of Agriculture of China, College of Animal Science and Technology, Northwest A&F UniversityAbstract Background Spermatogonial stem cells (SSCs) are the cornerstone of sperm production and thus perpetual male fertility. In clinics, transplantation of patient’s own SSCs into testes is a promising technique to restore fertility when male germ cells have been depleted by gonadotoxic therapies. Auto-transplantation of genetically modified SSCs even has the potential to treat male infertility caused by genetic mutations. However, SSCs are refractory to transfection approaches. Poly(amidoamine) (PAMAM) dendrimers have the unique three-dimensional architecture, surface charge, and high density of surface groups that are suitable for ligand attachment, thereby facilitating target delivery. The goal of this study was to elucidate whether PAMAM dendrimers can efficiently deliver short interfering RNAs (siRNAs) to SSCs. Methods and results We introduced cyclic arginine-glycine-aspartic acid (cRGD) peptides to the fifth generation of PAMAM dendrimers (G5) to generate PAMAM-cRGD dendrimers (G5-cRGD). The characterization of G5-cRGD was detected by Fourier transform infrared spectroscope (FTIR), transmission electron microscope (TEM), and the Cell Counting Kit-8 (CCK-8) assay. Confocal microscopy and flow cytometry were used to evaluate the delivery efficiency of siRNA by G5-cRGD to SSCs. The results showed that G5-cRGD encompassing siRNA could self-assemble into spherical structures with nanoscale size and possess high transfection efficiency, excellent endosomal escape ability, and low cytotoxicity, superior to a commercial transfection reagent Lipofectamine® 2000. Moreover, we demonstrated that G5-cRGD efficiently delivered siRNAs and triggered gene silencing. Conclusions This study thus provides a promising nanovector for siRNA delivery in SSCs, facilitating the future clinical application of SSC auto-transplantation with genetically modified cells with a hope to cure male infertility that is caused by genetic disorders.https://doi.org/10.1186/s13287-019-1506-4PAMAM-cRGDsiRNA deliveryNanoparticleSpermatogonial stem cells |
| spellingShingle | Tianjiao Li Qiwen Chen Yi Zheng Pengfei Zhang Xiaoxu Chen Junna Lu Yinghua Lv Shiguo Sun Wenxian Zeng PAMAM-cRGD mediating efficient siRNA delivery to spermatogonial stem cells Stem Cell Research & Therapy PAMAM-cRGD siRNA delivery Nanoparticle Spermatogonial stem cells |
| title | PAMAM-cRGD mediating efficient siRNA delivery to spermatogonial stem cells |
| title_full | PAMAM-cRGD mediating efficient siRNA delivery to spermatogonial stem cells |
| title_fullStr | PAMAM-cRGD mediating efficient siRNA delivery to spermatogonial stem cells |
| title_full_unstemmed | PAMAM-cRGD mediating efficient siRNA delivery to spermatogonial stem cells |
| title_short | PAMAM-cRGD mediating efficient siRNA delivery to spermatogonial stem cells |
| title_sort | pamam crgd mediating efficient sirna delivery to spermatogonial stem cells |
| topic | PAMAM-cRGD siRNA delivery Nanoparticle Spermatogonial stem cells |
| url | https://doi.org/10.1186/s13287-019-1506-4 |
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