Induced Pluripotent Stem Cells: Reprogramming Platforms and Applications in Cell Replacement Therapy

Abstract

The generation of induced pluripotent stem cells (iPSCs) from differentiated mature cells is one of the most promising technologies in the field of regenerative medicine. The ability to generate patient-specific iPSCs offers an invaluable reservoir of pluripotent cells, which could be genetically engineered and differentiated into target cells to treat various genetic and degenerative diseases once transplanted, hence counteracting the risk of graft versus host disease. In this context, we review the scientific research streams that lead to the emergence of iPSCs, the roles of reprogramming factors in reprogramming to pluripotency, and the reprogramming strategies. As iPSCs serve tremendous correction potentials for various diseases, we highlight the successes and challenges of iPSCs in cell replacement therapy and the synergy of iPSCs and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 gene editing tools in therapeutics research.

Disease modeling and stem cell immunoengineering in regenerative medicine using CRISPR/Cas9 systems

“…Development of CRISPR/Cas9 genome editing technology has enabled therapeutic application of CAR-T therapy through targeted intervention of endogenous genes [248] . Engineered CAR-T cells called iCARTs can be generated from patient-derived iPSCs with HLA-independent customizable antigen recognition, and they have been shown to kill cancer cells [248] , [249] , [250] . Together, these studies show that patient-derived iPSCs or hESCs used to generate NK cells in combination with CRISPR/Cas9-mediated gene modification could be an efficient cancer immunotherapy.…”

Modelling Mitochondrial Disease in Human Pluripotent Stem Cells: What Have We Learned?

“…Reprogramming to an iPSC fate involves reverting a terminally differentiated cell type, typically fibroblasts or peripheral blood mononuclear cells (PBMCs), to an uncommitted pluripotent stem cell fate by transient expression of Oct4 (Pou5f1), Sox2, Klf4, and c-Myc [ 67 ]. This can be accomplished using a range of technologies and Figure 3 outlines the technologies used to generate the mitochondrial stem cell models published to date [ 68 ].…”

“…Since the first extensive characterisations of iPSC mitochondrial disease models in 2013 [ 21 , 45 , 69 ], the choice of reprogramming method has largely been influenced by the predominant technology at the time of generation [ 68 ]. Integrative viral vectors like retrovirus and lentivirus were common in early iPSC studies due to their high efficiency and ease of use.…”

Utilising Induced Pluripotent Stem Cells in Neurodegenerative Disease Research: Focus on Glia

“…Currently, the most commonly used method to generate iPSCs is via the non-integrating Sendai viral vector [ 39 ]. There have also been many advances in iPSC generation and methods such as epigenetic regulation, microRNA manipulation, and nonviral methods have arisen in recent years [ 40 , 41 ]. iPSCs can be differentiated into various cell types, including neurons and glia, making them a powerful tool to study human CNS cells in vitro ( Figure 2 ).…”