A Cationic Stearamide‐based Solid Lipid Nanoparticle for Delivering Yamanaka Factors: Evaluation of the Transfection Efficiency
Abstract Induced pluripotent stem cells (IPSC) are preferred as an alternative source for regenerative medicine, disease modeling, and drug screening due to their unique properties. As seen from the previous studies in the literature, most of the vector systems to transfer reprogramming factors are...
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| Format: | Article |
| Language: | English |
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Wiley-VCH
2020-11-01
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| Series: | ChemistryOpen |
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| Online Access: | https://doi.org/10.1002/open.202000244 |
| _version_ | 1818027869329162240 |
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| author | Funda Alkan Hanife Sevgi Varlı Ph.D. Murat Demirbilek Ph.D. Engin Kaplan Ph.D. Nelisa Türkoğlu Laçin |
| author_facet | Funda Alkan Hanife Sevgi Varlı Ph.D. Murat Demirbilek Ph.D. Engin Kaplan Ph.D. Nelisa Türkoğlu Laçin |
| author_sort | Funda Alkan |
| collection | DOAJ |
| description | Abstract Induced pluripotent stem cells (IPSC) are preferred as an alternative source for regenerative medicine, disease modeling, and drug screening due to their unique properties. As seen from the previous studies in the literature, most of the vector systems to transfer reprogramming factors are viral‐based and have some well‐known limitations. This study aims to develop a non‐viral vector system for the transfection of reprogramming factors. Cationic stearamide lipid nanoparticles (CSLN) were prepared via the solvent diffusion method. The obtained CSLNs were used for the delivery of plasmid DNA (pDNA) encoding Oct3/4, Sox2, Klf4, and GFP to fibroblast cell lines. The optimization studies, for zeta potential and particle size of the conjugate, was performed to achieve high cell viability. CSLN63 with 36.5±0.06 mV zeta potential and 173.6±13.91 nm size was used for the transfection of Fibroblast cells. The transfection efficiency was observed by following GFP expression and was found as 70 %±0.11. The expression of the Oct4, Sox2, Klf4 was determined by RT‐qPCR; an increase was observed after the 12th cycle in Klf4 (Ct averages: 13,41), Sox2 (Ct averages; 12,4), Oct4 (Ct average; 13,77). The tendency of colonization was observed. The upregulation efficiency of Oct4 and SSEA‐1 with CSLN and another non‐viral vector designed for the transportation of Yamanaka factors developed in our lab previously were compared with flow cytometer analysis. |
| first_indexed | 2024-12-10T04:54:45Z |
| format | Article |
| id | doaj.art-5e02367a36cd420a862c22ae28b9d1a7 |
| institution | Directory Open Access Journal |
| issn | 2191-1363 |
| language | English |
| last_indexed | 2024-12-10T04:54:45Z |
| publishDate | 2020-11-01 |
| publisher | Wiley-VCH |
| record_format | Article |
| series | ChemistryOpen |
| spelling | doaj.art-5e02367a36cd420a862c22ae28b9d1a72022-12-22T02:01:31ZengWiley-VCHChemistryOpen2191-13632020-11-019111181118910.1002/open.202000244A Cationic Stearamide‐based Solid Lipid Nanoparticle for Delivering Yamanaka Factors: Evaluation of the Transfection EfficiencyFunda Alkan0Hanife Sevgi Varlı1Ph.D. Murat Demirbilek2Ph.D. Engin Kaplan3Ph.D. Nelisa Türkoğlu Laçin4Yıldız Technical University Molecular Biology and Genetic Department Istanbul 34220Yıldız Technical University Molecular Biology and Genetic Department Istanbul 34220Hacettepe University Advanced Technologies Application and Research Center Beytepe Ankara 06800 TurkeyBülent Ecevit University Faculty of Pharmacy Zonguldak TurkeyYıldız Technical University Molecular Biology and Genetic Department Istanbul 34220Abstract Induced pluripotent stem cells (IPSC) are preferred as an alternative source for regenerative medicine, disease modeling, and drug screening due to their unique properties. As seen from the previous studies in the literature, most of the vector systems to transfer reprogramming factors are viral‐based and have some well‐known limitations. This study aims to develop a non‐viral vector system for the transfection of reprogramming factors. Cationic stearamide lipid nanoparticles (CSLN) were prepared via the solvent diffusion method. The obtained CSLNs were used for the delivery of plasmid DNA (pDNA) encoding Oct3/4, Sox2, Klf4, and GFP to fibroblast cell lines. The optimization studies, for zeta potential and particle size of the conjugate, was performed to achieve high cell viability. CSLN63 with 36.5±0.06 mV zeta potential and 173.6±13.91 nm size was used for the transfection of Fibroblast cells. The transfection efficiency was observed by following GFP expression and was found as 70 %±0.11. The expression of the Oct4, Sox2, Klf4 was determined by RT‐qPCR; an increase was observed after the 12th cycle in Klf4 (Ct averages: 13,41), Sox2 (Ct averages; 12,4), Oct4 (Ct average; 13,77). The tendency of colonization was observed. The upregulation efficiency of Oct4 and SSEA‐1 with CSLN and another non‐viral vector designed for the transportation of Yamanaka factors developed in our lab previously were compared with flow cytometer analysis.https://doi.org/10.1002/open.202000244cationic nanoparticlefibroblastinduced pluripotent stem cellreprogramming factorsstearamide |
| spellingShingle | Funda Alkan Hanife Sevgi Varlı Ph.D. Murat Demirbilek Ph.D. Engin Kaplan Ph.D. Nelisa Türkoğlu Laçin A Cationic Stearamide‐based Solid Lipid Nanoparticle for Delivering Yamanaka Factors: Evaluation of the Transfection Efficiency ChemistryOpen cationic nanoparticle fibroblast induced pluripotent stem cell reprogramming factors stearamide |
| title | A Cationic Stearamide‐based Solid Lipid Nanoparticle for Delivering Yamanaka Factors: Evaluation of the Transfection Efficiency |
| title_full | A Cationic Stearamide‐based Solid Lipid Nanoparticle for Delivering Yamanaka Factors: Evaluation of the Transfection Efficiency |
| title_fullStr | A Cationic Stearamide‐based Solid Lipid Nanoparticle for Delivering Yamanaka Factors: Evaluation of the Transfection Efficiency |
| title_full_unstemmed | A Cationic Stearamide‐based Solid Lipid Nanoparticle for Delivering Yamanaka Factors: Evaluation of the Transfection Efficiency |
| title_short | A Cationic Stearamide‐based Solid Lipid Nanoparticle for Delivering Yamanaka Factors: Evaluation of the Transfection Efficiency |
| title_sort | cationic stearamide based solid lipid nanoparticle for delivering yamanaka factors evaluation of the transfection efficiency |
| topic | cationic nanoparticle fibroblast induced pluripotent stem cell reprogramming factors stearamide |
| url | https://doi.org/10.1002/open.202000244 |
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