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Improved therapeutic consistency and efficacy of CD317+ MSCs through stabilizing TSG6 by PTX3
Shaoquan Shi1,2, Si Chen3, Bowei Liang3
1Shenzhen Key Laboratory of Reproductive Immunology for Peri-Implantation, Shenzhen Zhongshan Institute for Reproduction and Genetics, Shenzhen Zhongshan Obstetrics & Gynecology Hospital, Fuqiang Avenue 1001, Shenzhen, 518060, Guangdong, People's Republic of China.
Background:
Previously, we have demonstrated that the batch variations of human platelet lysate (conventional MSC expansion medium) induce MSC heterogeneity and therapeutic inconsistency. On the other hand, the MSCs expanded with chemical defined medium have improved therapeutic consistency.
Methods:
In the current study, we studied the MSC subpopulation composition and variation in different types and batches of MSC expansion medium with scRNA-seq analysis.
Results:
MSCs expanded with different batches of media have higher levels of heterogeneity from the perspective of cell subpopulation composition at transcriptome levels and therapeutic inconsistency. The CD317+ subpopulation has enhanced immune suppression activities. And the percentage of CD317+ MSCs within MSCs is tightly correlated with its immune suppression activities, and also contributes to the heterogeneity and therapeutic inconsistency of MSCs. the CD317+ MSCs have increased expression levels of PTX3, which might stabilize the TSG6 protein and improve the therapeutic effects CONCLUSIONS: Thus, purifying CD317+ MSCs is one efficient strategy to reduce MSC heterogeneity and increase the therapeutic consistency of MSCs.
Insights
Batch variations in Mesenchymal Stem Cell (MSC) expansion media cause inconsistency. Purifying CD317+ MSCs reduces this heterogeneity, improving therapeutic outcomes.
Area of Science:
- Stem cell biology
- Regenerative medicine
Background:
- Batch variations in human platelet lysate (conventional MSC expansion medium) induce Mesenchymal Stem Cell (MSC) heterogeneity and therapeutic inconsistency.
- MSCs expanded in chemical defined media show improved therapeutic consistency.
Purpose of the Study:
- To investigate MSC subpopulation composition and variation across different MSC expansion media types and batches.
- To identify factors contributing to MSC heterogeneity and therapeutic inconsistency.
Main Methods:
- Single-cell RNA sequencing (scRNA-seq) analysis was employed to study MSC subpopulation composition.
- Comparison of MSCs expanded in different types and batches of MSC expansion media.
Main Results:
- MSCs expanded with different media batches exhibited higher heterogeneity at the transcriptome level and therapeutic inconsistency.
- A specific subpopulation, CD317+, demonstrated enhanced immune suppression activities.
- The percentage of CD317+ MSCs correlated with immune suppression and contributed to heterogeneity and inconsistency.
Conclusions:
- Purifying CD317+ MSCs is an effective strategy to reduce MSC heterogeneity.
- This purification enhances the therapeutic consistency of MSCs.
- CD317+ MSCs show increased PTX3 expression, potentially stabilizing TSG6 protein for improved therapeutic effects.
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