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Updated: Jan 20, 2026
Author Spotlight: Automating iPSC Culture for Enhanced Reproducibility
Published on: January 26, 2024
Engineered LINC MIR503HG-loaded extracellular vesicles maintain stemness and pluripotency during long-term hiPSCs
Jiaqi Zhu1,2, Xiaoren Zhu3,4, Wei Feng2,5
1Department of Hepatobiliary and Pancreatic Surgery, Affiliated Hospital of Nantong University, Medical School of Nantong University, Nantong, China.
Insights
Researchers developed MIR503HG-EVs to maintain stemness in human induced pluripotent stem cells (hiPSCs) during prolonged culture. This novel approach preserves hiPSC pluripotency and differentiation potential, crucial for regenerative medicine applications.
Area of Science:
- Stem Cell Biology
- Regenerative Medicine
- Molecular Biology
Background:
- Prolonged culture of human induced pluripotent stem cells (hiPSCs) leads to reduced stemness and pluripotency, hindering regenerative medicine.
- Maintaining hiPSC quality over extended periods is a significant challenge.
Purpose of the Study:
- To identify factors maintaining hiPSC stemness during prolonged culture.
- To develop an efficient delivery system for these factors to optimize hiPSC culture.
- To evaluate the impact of this system on hiPSC quality and differentiation potential.
Main Methods:
- Identification of LINC MIR503HG as a key factor for hiPSC stemness.
- Development of adipose-derived stem cell-derived extracellular vesicles (ADSC-EVs) for MIR503HG delivery (MIR503HG-EVs).
- Assessment of hiPSC morphology, viability, gene/protein expression (OCT4, SOX2, NANOG, MYC), chromosomal integrity, and differentiation potential after MIR503HG-EV treatment.
- Mechanistic studies involving AHCTF1 binding and nucleocytoplasmic transport of MYC mRNA.
Main Results:
- MIR503HG-EV treatment maintained hiPSC compact morphology, viability, and expression of pluripotency markers (OCT4, SOX2, NANOG).
- Chromosomal integrity was preserved, with no detected anomalies.
- MIR503HG facilitated MYC mRNA transport, boosting MYC protein production and activating stemness pathways.
- MIR503HG-EVs improved differentiation efficiency of high-passage hiPSCs into definitive endoderm, pancreatic, and hepatic lineages.
Conclusions:
- MIR503HG-EVs represent a convenient, efficient, and safe method for maintaining high-passage hiPSCs.
- This approach overcomes challenges associated with prolonged hiPSC culture, enhancing their utility in regenerative medicine.
- The study elucidates a novel mechanism involving MIR503HG, AHCTF1, and MYC in maintaining stemness.
Abstract:
The reduction of stemness and pluripotency during the prolonged culture of hiPSCs poses a significant challenge in regenerative medicine. This study identified LINC MIR503HG as a factor involved in maintaining hiPSCs stemness. Consequently, it developed a highly efficient delivery system based on ADSC-EVs, named MIR503HG-EVs, to optimize the culture strategy for hiPSCs. During the extended culture, MIR503HG-EV-treated hiPSCs developed into colonies with more compact morphology, an increased percentage of viable cells, as well as elevated OCT4, SOX2, and NANOG expression. Furthermore, these cells maintained their chromosomal integrity, as no karyotypic anomalies were detected. Mechanistic studies demonstrated that MIR503HG selectively bound AHCTF1 to facilitate the active nucleocytoplasmic transport of MYC mRNA. This resulted in significantly augmented MYC protein production, which activated the stemness regulatory network. Concurrently, MIR503HG-EVs mitigated the decline in differentiation potential of hiPSCs after several passages by modulating the chromatin accessibility of stemness transcription factors and modifying energy metabolism, including glycolysis and oxidative phosphorylation pathways. Moreover, treatment with MIR503HG-EVs significantly enhanced the differentiation efficiency of high-passage hiPSCs into definitive endoderm, pancreatic, and hepatic lineages, thereby achieving a level of proficiency comparable to that of low-passage clones. Overall, this study identified the addition of MIR503HG-EVs as a convenient, efficient, and safe approach for maintaining high-passage hiPSCs.
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