iPSCs
endogenous pluripotency program and the conversion of the somatic cells into iPSCs. The efficiency of protein-induced iPSC generation is influenced by various factors, such as the quality and purity of the reprogramming proteins, the delivery method, and the cell type being reprogrammed. The mechanistic understanding of how these proteins interact with their target genes and trigger the complex molecular events that result in iPSC generation is still an active area of research. 43
Benefits and drawbacks
Generating induced pluripotent stem cells (iPSCs) via protein delivery is considered better than other methods, such as viral vectors, because it avoids genetic modification and reduces the risk of tumorigenicity, making it safer for clinical applications. 44 The protein transduction method eliminates issues like insertional mutagenesis, residual expression and re-activation of reprogramming factors, uncontrolled silencing of transgenes, apoptosis, cell senescence, and strong immunogenicity, which are associated with virus-based reprogramming. Additionally, protein delivery methods can be more efficient and provide a higher yield of specific cell types, such as A9-subtype ventral midbrain dopamine neurons. Overall, protein delivery methods offer a safer and more controlled approach to iPSC generation, which is crucial for their use in regenerative medicine and disease modelling. 45 The use of protein delivery to generate induced pluripotent stem cells (iPSCs) has some drawbacks. Firstly, protein-based reprogramming methods are generally less efficient than other methods, such as viral vectors. 46 This is due to the slow movement of proteins. Protein-based strategies have slower kinetics compared to other methods, which can be a disadvantage in turning them into iPSCs as it takes longer for the proteins to enter the cell. Secondly, the recombinant proteins used in these approaches can be challenging to reproducibly purify in sufficient quantities and with the required quality. Furthermore, multiple transfections of proteins may be needed which can be time-consuming and labour-intensive. 47 Finally, protein-based approaches may lead to the formation of inclusion bodies, which can negatively impact the effectiveness of iPSC generation as these cells may not be useable.
43 Hu K. (2014). All roads lead to induced pluripotent stem cells: the technologies of iPSC generation. Stem cells and development , 23 (12), 1285 – 1300. 44 Vijakumaran, U., Nordin, F., Abdul Hamid, Z., Abdullah, M., & Gee, J.T. (2022). TAT Κappa (TATΚ): A Novel Cell Penetrating Peptide for Delivery of Pluripotent Proteins into Target Cells. Sains Malaysiana . 45 Hu K. (2014). All roads lead to induced pluripotent stem cells: the technologies of iPSC generation. Stem cells and development , 23 (12), 1285 – 1300. 46 Wang, Q., Vossen, A.M., Ikeda, Y., & Devaux, P. (2019). Measles vector as a multigene delivery platform facilitating iPSC reprogramming. Gene Therapy, 26 , 151-164. 47 Vijakumaran, U., Nordin, F., Abdul Hamid, Z., Abdullah, M., & Gee, J.T. (2022). TAT Κappa (TATΚ): A Novel Cell Penetrating Peptide for Delivery of Pluripotent Proteins into Target Cells. Sains Malaysiana .
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