A virus-free vector for the transfection of somatic cells to obtain IPSC

Varlı, Hanife Sevgi; Alkan, Funda; Demirbilek, Murat; Türkoğlu, Nelisa

doi:10.1007/s11051-019-4668-1

Cited by 12 publications

(12 citation statements)

References 35 publications

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“…The first one is the stearamide nanoparticle (CSLN), which was designed and synthesized in the study. The second system is octadecylamine nanoparticle (OLN), which has been synthesized in our previous studies [22] …”

Section: Methodsmentioning

confidence: 99%

“…The second system is octadecylamine nanoparticle (OLN), which has been synthesized in our previous studies. [22] The stearamide nanoparticle was synthesized by the solvent diffusion method, and parameters were optimized to obtain convenient particle size and zeta potential. Briefly, 0.75 M stearamide was dissolved in 4 mL of chloroform-ethanol (1 : 1) solvent system to obtain a stearamide solution as a lipid phase.…”

Section: Solid Lipid Nanoparticle Synthesismentioning

confidence: 99%

“…Cell cytotoxicity of the SLNP system was determined on the mouse fibroblast‐like cell line (L929). The octadecylamine‐based lipid nanoparticle (OLN) was prepared in our previous studies, [22] and the two novel nanocarrier systems for cell reprogramming were compared in this study.…”

Section: Introductionmentioning

confidence: 99%

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A Cationic Stearamide‐based Solid Lipid Nanoparticle for Delivering Yamanaka Factors: Evaluation of the Transfection Efficiency

et al. 2020

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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.

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Section: Methodsmentioning

confidence: 99%

Section: Solid Lipid Nanoparticle Synthesismentioning

confidence: 99%

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A Cationic Stearamide‐based Solid Lipid Nanoparticle for Delivering Yamanaka Factors: Evaluation of the Transfection Efficiency

et al. 2020

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“…Arginine-terminated generation 4 polyamidoamine (G4Arg) nanoparticles were successfully used for the delivery of a single plasmid construct carrying the four transcription factors into mouse embryonic fibroblasts to generate iPSCs [ 93 ]. Recently, several other types of nanoparticles were used for the reprograming of somatic cells to iPSCs including octadecylamine-based cationic lipid nanoparticles [ 9 , 94 ]. In addition, protein and peptide-based systems were used to reprogram somatic cells to iPSCs [ 95 , 96 , 97 , 98 ].…”

Section: Nanoparticles In Ipsc Generation and Precision Medicinementioning

confidence: 99%

Transgene Delivery to Human Induced Pluripotent Stem Cells Using Nanoparticles

2021

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Human induced pluripotent stem cells (HiPSCs) and HiPSCs-derived cells have the potential to revolutionize regenerative and precision medicine. Genetically reprograming somatic cells to generate HiPSCs and genetic modification of HiPSCs are considered the key procedures for the study and application of HiPSCs. However, there are significant technical challenges for transgene delivery into somatic cells and HiPSCs since these cells are known to be difficult to transfect. The existing methods, such as viral transduction and chemical transfection, may introduce significant alternations to hiPSC culture which affect the potency, purity, consistency, safety, and functional capacity of HiPSCs. Therefore, generation and genetic modification of HiPSCs through non-viral approaches are necessary and desirable. Nanotechnology has revolutionized fields from astrophysics to biology over the past two decades. Increasingly, nanoparticles have been used in biomedicine as powerful tools for transgene and drug delivery, imaging, diagnostics, and therapeutics. The most successful example is the recent development of SARS-CoV-2 vaccines at warp speed to combat the 2019 coronavirus disease (COVID-19), which brought nanoparticles to the center stage of biomedicine and demonstrated the efficient nanoparticle-mediated transgene delivery into human body. Nanoparticles have the potential to facilitate the transgene delivery into the HiPSCs and offer a simple and robust approach. Nanoparticle-mediated transgene delivery has significant advantages over other methods, such as high efficiency, low cytotoxicity, biodegradability, low cost, directional and distal controllability, efficient in vivo applications, and lack of immune responses. Our recent study using magnetic nanoparticles for transfection of HiPSCs provided an example of the successful applications, supporting the potential roles of nanoparticles in hiPSC biology. This review discusses the principle, applications, and significance of nanoparticles in the transgene delivery to HiPSCs and their successful application in the development of COVID-19 vaccines.

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“…Varlı ve ark. lipit bazlı bir nano-taşıyıcı sistemi dizayn ederek ilk kez fare fibroblast hücrelerini yeniden programlanması amacı ile kullanmışlardır 66 .…”

Section: Tablo I İpkh Elde Edilmesi Için Kullanılan Vektör Sistemleri Ve Verimlilikleriunclassified

İndüklenmiş Pluripotent Kök Hücrelerin Elde Edilmesi ve Rejeneratif Tıpta Uygulanabilirliği

Cesur

Türkoğlu

2021

Uludağ Üniversitesi Tıp Fakültesi Dergisi

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ÖZET2006 yılında Takahashi ve Yamanaka dört transkripsiyon faktörünün (Oct4, Sox2, Klf4 ve c-Myc) fibroblast hücrelerine aktarılması ve bu transkripsiyon faktörlerinin ifadesinin pluripotent kök hücre elde etmek için yeterli olduğunu bildirmiş ve somatik hücrelerin geriye programlanarak elde edilen bu hücreler indüklenmiş pluripotent kök hücreler (İPKH) olarak adlandırılmıştır. Daha sonraki yıllarda transkripsiyon faktörleri ve yeniden programlama şartlarının optimizasyonu ile ilgili birçok çalışma yapılmıştır. Bugüne kadar farklı somatik hücrelere transkripsiyon faktörlerinin farklı metotları ile tanıtımı ya da transkripsiyon faktörlerinin farklı kombinasyonlarının kullanımının etkisi araştırma konusu olmuştur. Somatik hücrelerin yeniden programlanması amacı ile birçok farklı vektör sistemi bulunmaktadır. Bu vektör çeşitlerinin İPKH eldesi için verimlilikleri birbirlerinden farklılık göstermektedir. Bu derlemede, kök hücrelerin genel özellikleri ve uygulama alanlarının irdelenmesinin yanı sıra ağırlıklı olarak indüklenmiş pluripotent kök hücrelerinin elde edilmesi üzerinde durulmuştur. Ayrıca İPKH'lerin klinik amaçlı kullanım potansiyellerine de değinilmektedir. Anahtar Kelimeler: Kök hücreler. Kök hücre çeşitleri. Rejeneratif tıp. İndüklenmiş pluripotent kök hücreler. İndüklenmiş kök hücre eldesi.

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A virus-free vector for the transfection of somatic cells to obtain IPSC

Cited by 12 publications

References 35 publications

A Cationic Stearamide‐based Solid Lipid Nanoparticle for Delivering Yamanaka Factors: Evaluation of the Transfection Efficiency

A Cationic Stearamide‐based Solid Lipid Nanoparticle for Delivering Yamanaka Factors: Evaluation of the Transfection Efficiency

Transgene Delivery to Human Induced Pluripotent Stem Cells Using Nanoparticles

İndüklenmiş Pluripotent Kök Hücrelerin Elde Edilmesi ve Rejeneratif Tıpta Uygulanabilirliği

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