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Characterization of Senescence of Human Adipose-Derived Stem Cells After Long-Term Expansion

  • Nhat Chau Truong
  • Khanh Hong-Thien Bui
  • Phuc Van PhamEmail author
Conference paper
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 1084)

Abstract

Introduction: Since the 1980s, adipose-derived stem cells (ASCs) have become a powerful and potential source for stem cell-based therapy, regenerative medicine, and even drug delivery in cancer treatment. The development of off-the-shelf mesenchymal stem cells (MSCs), including ASCs, has rapidly advanced in recent years with several clinical trials and approved products. In this technology, ASCs should be expanded long term in order to harvest higher cell number. In this study, senescence of ASCs after long-term expansion was evaluated. Methods: Human ASCs (hASCs) were isolated and cultured continuously at a density of 103 cells/cm2 up to passage 15. The cells were assessed for aging via changes in the following: characteristics of MSCs, mitochondrial activity, accumulation of beta-galactosidase, and expression of tumor suppressor genes. Results: The results showed that following in vitro expansion to the 15th passage, ASCs did not show changes in immunophenotype, except for decreased expression of CD105. However, the cells increased in size and in shape and complexity (toward the “fried egg” morphology). They also almost ceased to proliferate in passage 15. Nonetheless, they maintained in vitro differentiation potential toward osteoblasts, chondrocytes, and adipocytes. Expression of tumor suppressor genes p53 and p16 did not significantly change, while p27 was significantly downregulated. Mitochondrial activities also decreased slightly in culture from passage 5 to passage 10 and remained stable to passage 15. ASCs also showed increased accumulation of beta-galactosidase in culture, but it was negligible. Conclusion: In conclusion, hASCs exhibited some particular characteristics of aged stem cells when the number of subculture cells increased. However, up to passage 10, ASCs also retained almost all of the characteristics of MSCs.

Keywords

Aging Senescence Long-term In vitro culture Mesenchymal stem cells Adipose-derived stem cells 

Abbreviations

ASCs:

Adipose-derived stem cells

BM-MSCs:

Bone marrow-derived mesenchymal stem cells

CFU-F:

Fibroblast colony-forming units

DE:

Germany

FBS:

Fetal bovine serum

FCM:

Flow cytometry

FSC:

Forward scatter

hASCs:

Human adipose-derived stem cells

iPSCs:

Induced pluripotent stem cells

KR:

Korean

MSCs:

Mesenchymal stem cells

PBS:

Phosphate-buffered saline

ROS:

Reactive oxygen species

SA-β-gal:

Senescence-associated β-galactosidase enzyme

SSC:

Side scatter

SVF:

Stromal vascular fraction

UK:

United Kingdom

USA:

United States

Notes

Acknowledgment

This research was supported by Viet Nam National University Ho Chi Minh City via projects Grant No. C2016-18-18 and TX2017-18-02 and by VNUHCM-University of Science via project Grant No. T2017-43.

Authors’ Contributions

NCT, PVP, and KHTB designed the study and read and corrected the manuscript. NCT wrote the manuscript, proliferated hASC samples, and evaluated gene expression, mitochondrial activity, and beta-galactosidase accumulation. NCT, PVP, and KHTB evaluated mesenchymal characteristics. All authors read and approved the final manuscript.

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Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  • Nhat Chau Truong
    • 1
    • 2
  • Khanh Hong-Thien Bui
    • 3
  • Phuc Van Pham
    • 1
    • 2
    Email author
  1. 1.Laboratory of Stem Cell Research and ApplicationVNUHCM University of ScienceHo Chi Minh CityVietnam
  2. 2.Stem Cell InstituteVNUHCM University of ScienceHo Chi Minh CityVietnam
  3. 3.University Medical CenterUniversity of Medicine and PharmacyHo Chi Minh CityVietnam

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