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Peer-reviewedCC BY 4.0Cells

hiPSC Extract Attenuates Senescence-Associated Phenotypes in AB-MSCs In Vitro

  • Yuanyuan Pan1
  • Lumiao Zhang2
  • Rongyuan Zhu2
  • Fangfang Sun3
  • Guofeng Wu2,3
  1. 1Department of the First Outpatient, Nanjing Stomatological Hospital, Affiliated Hospital of Medical School, Institute of Stomatology, Nanjing University, Nanjing 210000, China
  2. 2Department of Prosthodontics, Nanjing Stomatological Hospital, Affiliated Hospital of Medical School, Institute of Stomatology, Nanjing University, Nanjing 210000, China
  3. 3Digital Engineering Center of Stomatology, Nanjing Stomatological Hospital, Affiliated Hospital of Medical School, Institute of Stomatology, Nanjing University, Nanjing 210000, China
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Abstract

Background: Bone marrow mesenchymal stromal cells (BMSCs) are widely used as seed cells in bone tissue engineering. However, sufficient cell quantities for effective engineering typically require prolonged in vitro expansion of BMSCs, which can lead to cellular senescence after excessive subculturing. Objective: This study aimed to investigate the effects of human induced pluripotent stem cell (hiPSC) extract on replicative senescence, clonogenic ability, proliferation-associated characteristics, and osteogenic differentiation capacity of human alveolar bone-derived mesenchymal stromal cells (AB-MSCs). Methods: AB-MSCs were isolated from alveolar bone fragments discarded during dental implant surgery. Cells were divided into three experimental groups: P3 AB-MSCs (Young), P10 AB-MSCs (senescence-associated, YRS), and P10 AB-MSCs treated with hiPSC extract (YRSiE). Senescence markers, clonogenic ability, proliferation-associated characteristics, osteogenic potential, mRNA sequencing and pathway-related expression changes were compared among groups. Small RNA sequencing on hiPSCs and AB-MSCs with qRT-PCR validation were performed to identify candidate miRNAs in hiPSC extract that were differentially expressed in this setting. Results: In the presence of hiPSC extract, senescent AB-MSCs showed altered morphology, a lower proportion of β-galactosidase-positive cells, higher Ki67 expression, and lower expression of the senescence-associated genes and proteins P16 and P21. Treated senescent AB-MSCs also showed higher proliferation-associated metabolic activity, clonogenicity, and osteogenic differentiation than untreated senescent cells. Pluripotency-related genes (NANOG, OCT-4) and osteogenic markers (ALP, RUNX2, OCN) were higher in the treated group. qRT-PCR validation of small RNA sequencing data showed that miR-514a-3p, miR-582-5p and miR-520c-3p were upregulated in senescent AB-MSCs after hiPSC extract treatment. Concurrently, pathway enrichment and pathway-related expression changes were consistent with an association with the NF-κB pathway. Conclusion: In this in vitro study, hiPSC extract attenuates senescence-associated phenotypes in AB-MSCs, and the observed transcriptional and pathway-related expression changes were consistent with NF-κB pathway enrichment, with the identified miRNAs upregulated in treated cells and previously reported to target NF-κB pathway components. However, these mechanisms require further validation.

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Cite

BibTeXRIS
BibTeX
@article{pan2026hipsc,
  title = {{hiPSC Extract Attenuates Senescence-Associated Phenotypes in AB-MSCs In Vitro}},
  author = {Yuanyuan Pan and Lumiao Zhang and Rongyuan Zhu and Fangfang Sun and Guofeng Wu},
  journal = {Cells},
  year = {2026},
  month = oct,
  doi = {10.3390/cells15211965},
  url = {https://www.mdpi.com/2073-4409/15/21/1965},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - hiPSC Extract Attenuates Senescence-Associated Phenotypes in AB-MSCs In Vitro
AU  - Yuanyuan Pan
AU  - Lumiao Zhang
AU  - Rongyuan Zhu
AU  - Fangfang Sun
AU  - Guofeng Wu
PY  - 2026
DA  - 2026/10/28
JO  - Cells
DO  - 10.3390/cells15211965
UR  - https://www.mdpi.com/2073-4409/15/21/1965
AB  - Background: Bone marrow mesenchymal stromal cells (BMSCs) are widely used as seed cells in bone tissue engineering. However, sufficient cell quantities for effective engineering typically require prolonged in vitro expansion of BMSCs, which can lead to cellular senescence after excessive subculturing. Objective: This study aimed to investigate the effects of human induced pluripotent stem cell (hiPSC) extract on replicative senescence, clonogenic ability, proliferation-associated characteristics, and osteogenic differentiation capacity of human alveolar bone-derived mesenchymal stromal cells (AB-MSCs). Methods: AB-MSCs were isolated from alveolar bone fragments discarded during dental implant surgery. Cells were divided into three experimental groups: P3 AB-MSCs (Young), P10 AB-MSCs (senescence-associated, YRS), and P10 AB-MSCs treated with hiPSC extract (YRSiE). Senescence markers, clonogenic ability, proliferation-associated characteristics, osteogenic potential, mRNA sequencing and pathway-related expression changes were compared among groups. Small RNA sequencing on hiPSCs and AB-MSCs with qRT-PCR validation were performed to identify candidate miRNAs in hiPSC extract that were differentially expressed in this setting. Results: In the presence of hiPSC extract, senescent AB-MSCs showed altered morphology, a lower proportion of β-galactosidase-positive cells, higher Ki67 expression, and lower expression of the senescence-associated genes and proteins P16 and P21. Treated senescent AB-MSCs also showed higher proliferation-associated metabolic activity, clonogenicity, and osteogenic differentiation than untreated senescent cells. Pluripotency-related genes (NANOG, OCT-4) and osteogenic markers (ALP, RUNX2, OCN) were higher in the treated group. qRT-PCR validation of small RNA sequencing data showed that miR-514a-3p, miR-582-5p and miR-520c-3p were upregulated in senescent AB-MSCs after hiPSC extract treatment. Concurrently, pathway enrichment and pathway-related expression changes were consistent with an association with the NF-κB pathway. Conclusion: In this in vitro study, hiPSC extract attenuates senescence-associated phenotypes in AB-MSCs, and the observed transcriptional and pathway-related expression changes were consistent with NF-κB pathway enrichment, with the identified miRNAs upregulated in treated cells and previously reported to target NF-κB pathway components. However, these mechanisms require further validation.
SN  - 2073-4409
ER  -