mRNA-Lipid Nanoparticle-Mediated Reprogramming and Standard Sendai Virus Reprogramming: Generation of iPSCs and...

Area of Science:

• Stem Cell Biology
• Gene Therapy
• Regenerative Medicine

Background:

• Non-integrating Sendai virus vectors are the standard for induced pluripotent stem cell (iPSC) reprogramming.
• Current methods face challenges in cost, regulatory oversight, and scalability for regenerative medicine and biobanking.
• Traditional mRNA delivery methods like electroporation can be physiologically disruptive.

Purpose of the Study:

• To evaluate a novel mRNA-lipid nanoparticle (mRNA-LNP) platform for iPSC reprogramming.
• To compare the efficacy and characteristics of mRNA-LNP reprogrammed iPSCs against Sendai virus-reprogrammed iPSCs.
• To assess the potential of mRNA-LNPs for regenerative medicine and large-scale biobanking applications.

Main Methods:

• Reprogramming of peripheral blood mononuclear cells (PBMCs) from the same donor using both Sendai virus and mRNA-LNP methods.
• Analysis of genomic integrity, pluripotency markers, and differentiation capacity of resulting iPSCs.
• Differentiation of iPSCs into cardiomyocytes to assess functional and phenotypic recapitulation.

Main Results:

• mRNA-LNP reprogrammed iPSCs maintained genomic integrity and donor-specific variants (e.g., KCNH2 c.2398+5G>T).
• mRNA-LNP iPSCs exhibited characteristic pluripotency markers, morphology, and trilineage differentiation capacity comparable to Sendai virus iPSCs.
• mRNA-LNP reprogrammed iPSCs differentiated into functional cardiomyocytes with disease-specific phenotypes, achieved in significantly fewer passages.

Conclusions:

• The mRNA-LNP platform provides an effective, virus-independent alternative for iPSC reprogramming.
• This method significantly reduces reprogramming timelines and costs compared to Sendai virus vectors.
• mRNA-LNP reprogramming is a promising technology for advancing regenerative medicine, precision medicine, and scalable biobanking.