Salicylic diamines selectively eliminate residual undifferentiated cells from pluripotent stem cell-derived
Karsten Burkert1, Hadiseh Taheri1, Sarkawt Hamad1
1Center for Physiology and Pathophysiology, Institute for Neurophysiology, Medical Faculty, University of Cologne, Cologne, Germany.
Abstract:
Clinical translation of pluripotent stem cell (PSC) derivatives is hindered by the tumorigenic risk from residual undifferentiated cells. Here, we identified salicylic diamines as potent agents exhibiting toxicity to murine and human PSCs but not to cardiomyocytes (CMs) derived from them. Half maximal inhibitory concentrations (IC50) of small molecules SM2 and SM6 were, respectively, 9- and 18-fold higher for human than murine PSCs, while the IC50 of SM8 was comparable for both PSC groups. Treatment of murine embryoid bodies in suspension differentiation cultures with the most effective small molecule SM6 significantly reduced PSC and non-PSC contamination and enriched CM populations that would otherwise be eliminated in genetic selection approaches. All tested salicylic diamines exerted their toxicity by inhibiting the oxygen consumption rate (OCR) in PSCs. No or only minimal and reversible effects on OCR, sarcomeric integrity, DNA stability, apoptosis rate, ROS levels or beating frequency were observed in PSC-CMs, although effects on human PSC-CMs seemed to be more deleterious at higher SM-concentrations. Teratoma formation from SM6-treated murine PSC-CMs was abolished or delayed compared to untreated cells. We conclude that salicylic diamines represent promising compounds for PSC removal and enrichment of CMs without the need for other selection strategies.
Insights
Salicylic diamines effectively eliminate residual pluripotent stem cells (PSCs) while sparing derived cardiomyocytes (CMs). This approach enhances CM purity and reduces teratoma risk, aiding clinical translation.
Area of Science:
- Stem cell biology
- Cardiovascular research
- Drug discovery
Background:
- Clinical applications of pluripotent stem cells (PSCs) are limited by the risk of teratoma formation from residual undifferentiated cells.
- Enriching cardiomyocyte (CM) populations after differentiation is crucial for therapeutic use.
Purpose of the Study:
- To identify compounds that selectively eliminate undifferentiated PSCs without harming differentiated CMs.
- To evaluate the efficacy of salicylic diamines in PSC removal and CM enrichment.
Main Methods:
- Screening of salicylic diamines for selective toxicity against murine and human PSCs.
- Assessing the impact of compounds on PSC-CMs, including oxygen consumption rate (OCR), cellular integrity, and teratoma formation.
- Evaluating the effectiveness of SM6 in reducing PSC contamination in differentiation cultures.
Main Results:
- Salicylic diamines demonstrated potent toxicity to PSCs but not to derived CMs.
- SM6 significantly reduced PSC contamination and enriched CM populations in vitro.
- Salicylic diamines inhibited PSC OCR, with minimal reversible effects on CMs.
- Teratoma formation was abolished or delayed in PSC-CMs treated with SM6.
Conclusions:
- Salicylic diamines are promising agents for selective PSC removal and CM enrichment.
- These compounds offer a non-genetic strategy to improve the safety and efficacy of stem cell-derived therapies.
- Further investigation into salicylic diamines could facilitate the clinical translation of PSC-based regenerative medicine.
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