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Determining Osteogenic Differentiation Efficacy of Pluripotent Stem Cells by Telomerase Activity

  • Original Article
  • Published:
Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Background:

Bone tissue engineering based on pluripotent stem cells (PSCs) is a new approach to deal with bone defects. Protocols have been developed to generate osteoblasts from PSCs. However, the low efficiency of this process is still an important issue that needs to be resolved. Many studies have aimed to improve efficiency, but developing accurate methods to determine efficacy is also critical. Studies using pluripotency to estimate efficacy are rare. Telomerase is highly associated with pluripotency.

Methods:

We have described a quantitative method to measure telomerase activity, telomeric repeat elongation assay based on quartz crystal microbalance (QCM). To investigate whether this method could be used to determine the efficiency of in vitro osteogenic differentiation based on pluripotency, we measured the pluripotency pattern of cultures through stemness gene expression, proliferation ability and telomerase activity, measured by QCM.

Results:

We showed that the pluripotency pattern determined by QCM was similar to the patterns of proliferation ability and gene expression, which showed a slight upregulation at the late stages, within the context of the general downregulation tendency during differentiation. Additionally, a comprehensive gene expression pattern covering nearly every stage of differentiation was identified.

Conclusion:

Therefore, this assay may be powerful tools for determining the efficiency of differentiation systems based on pluripotency. In this study, we not only introduce a new method for determining efficiency based on pluripotency, but also provide more information about the characteristics of osteogenic differentiation which help facilitate future development of more efficient protocols.

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References

  1. Mateizel I, De Becker A, Van de Velde H, De Rycke M, Van Steirteghem A, Cornelissen R, et al. Efficient differentiation of human embryonic stem cells into a homogeneous population of osteoprogenitor-like cells. Reprod Biomed Online. 2008;16:741–53.

    Article  Google Scholar 

  2. Duplomb L, Dagouassat M, Jourdon P, Heymann D. Concise review: embryonic stem cells: a new tool to study osteoblast and osteoclast differentiation. Stem Cells. 2007;25:544–52.

    Article  CAS  Google Scholar 

  3. Phillips BW, Belmonte N, Vernochet C, Ailhaud G, Dani C. Compactin enhances osteogenesis in murine embryonic stem cells. Biochem Biophys Res Commun. 2001;284:478–84.

    Article  CAS  Google Scholar 

  4. Langenbach F, Handschel J. Effects of dexamethasone, ascorbic acid and β-glycerophosphate on the osteogenic differentiation of stem cells in vitro. Stem Cell Res Ther. 2013;4:117.

    Article  Google Scholar 

  5. Lou X. Induced pluripotent stem cells as a new strategy for osteogenesis and bone regeneration. Stem Cell Rev. 2015;11:645–51.

    Article  CAS  Google Scholar 

  6. Yamashita A, Nishikawa S, Rancourt DE. Microenvironment modulates osteogenic cell lineage commitment in differentiated embryonic stem cells. PLoS One. 2010;5:e9663.

    Article  Google Scholar 

  7. Kärner E, Bäckesjö CM, Cedervall J, Sugars RV, Ahrlund-Richter L, Wendel M. Dynamics of gene expression during bone matrix formation in osteogenic cultures derived from human embryonic stem cells in vitro. Biochim Biophys Acta. 2009;1790:110–8.

    Article  Google Scholar 

  8. Li F, Bronson S, Niyibizi C. Derivation of murine induced pluripotent stem cells (iPS) and assessment of their differentiation toward osteogenic lineage. J Cell Biochem. 2010;109:643–52.

    Article  CAS  Google Scholar 

  9. Tsukune N, Naito M, Kubota T, Ozawa Y, Nagao M, Ohashi A, et al. Lamin A overexpression promotes osteoblast differentiation and calcification in the MC3T3-E1 preosteoblastic cell line. Biochem Biophys Res Commun. 2017;488:664–70.

    Article  CAS  Google Scholar 

  10. Manokawinchoke J, Osathanon T, Egusa H, Pavasant P. Hypoxia enhances osteogenic differentiation in retinoic acid-treated murine-induced pluripotent stem cells. Tissue Eng Regen Med. 2016;13:547–53.

    Article  CAS  Google Scholar 

  11. Kim YD, Pofali P, Park TE, Singh B, Cho K, Maharjan S, et al. Gene therapy for bone tissue engineering. Tissue Eng Regen Med. 2016;13:111–25.

    Article  CAS  Google Scholar 

  12. Taiani JT, Krawetz RJ, Zur Nieden NI, Elizabeth Wu Y, Kallos MS, Matyas JR, et al. Reduced differentiation efficiency of murine embryonic stem cells in stirred suspension bioreactors. Stem Cells Dev. 2010;19:989–98.

    Article  CAS  Google Scholar 

  13. Greider CW, Blackburn EH. Identification of a specific telomere terminal transferase activity in tetrahymena extracts. Cell. 1985;43:405–13.

    Article  CAS  Google Scholar 

  14. Blackburn EH. Structure and function of telomeres. Nature. 1991;350:569–73.

    Article  CAS  Google Scholar 

  15. Buseman C, Wright W, Shay J. Is telomerase a viable target in cancer? Mutat Res. 2012;730:90–7.

    Article  CAS  Google Scholar 

  16. Wang J, Wu L, Ren J, Qu X. Visualizing human telomerase activity with primer-modified au nanoparticles. Small. 2012;8:259–64.

    Article  CAS  Google Scholar 

  17. Zhou Y, Zhou P, Xin Y, Wang J, Zhu Z, Hu J, et al. Trend of telomerase activity change during human iPSC self-renewal and differentiation revealed by a quartz crystal microbalance based assay. Sci Rep. 2014;4:6978.

    Article  CAS  Google Scholar 

  18. Wang J, Liu L, Ma H. Label-free real-time investigation of the effect of telomerase inhibitors based on quartz crystal microbalance measurement. Sens Actuators B Chem. 2017;239:943–50.

    Article  CAS  Google Scholar 

  19. Draper JS, Pigott C, Thomson JA, Andrews PW. Surface antigens of human embryonic stem cells: changes upon differentiation in culture. J Anat. 2002;200:249–58.

    Article  CAS  Google Scholar 

  20. Wang M, Deng Y, Zhou P, Luo Z, Li Q, Xie B, et al. In vitro culture and directed osteogenic differentiation of human pluripotent stem cells on peptides-decorated two-dimensional microenvironment. ACS Appl Mater Interfaces. 2015;7:4560–72.

    Article  CAS  Google Scholar 

  21. Wilkinson DG, Bhatt S, Herrmann BG. Expression pattern of the mouse T gene and its role in mesoderm formation. Nature. 1990;343:657–9.

    Article  CAS  Google Scholar 

  22. Deutsch U, Dressler GR, Gruss P. Pax 1, a member of a paired box homologous murine gene family, is expressed in segmented structures during development. Cell. 1988;53:617–25.

    Article  CAS  Google Scholar 

  23. Mankoo BS, Skuntz S, Harrigan I, Grigorieva E, Candia A, Wright CV, et al. The concerted action of Meox homeobox genes is required upstream of genetic pathways essential for the formation, patterning and differentiation of somites. Development. 2003;130:4655–64.

    Article  CAS  Google Scholar 

  24. Mori-Akiyama Y, Akiyama H, Rowitch DH, de Crombrugghe B. Sox9 is required for determination of the chondrogenic cell lineage in the cranial neural crest. Proc Natl Acad Sci U S A. 2003;100:9360–5.

    Article  CAS  Google Scholar 

  25. Nakashima K, Zhou X, Kunkel G, Zhang Z, Deng JM, Behringer RR, et al. The novel zinc finger-containing transcription factor osterix is required for osteoblast differentiation and bone formation. Cell. 2002;108:17–29.

    Article  CAS  Google Scholar 

  26. Nakashima K, de Crombrugghe B. Transcriptional mechanisms in osteoblast differentiation and bone formation. Trends Genet. 2003;19:458–66.

    Article  CAS  Google Scholar 

  27. Woll NL, Heaney JD, Bronson SK. Osteogenic nodule formation from single embryonic stem cell-derived progenitors. Stem Cells Dev. 2006;15:865–79.

    Article  CAS  Google Scholar 

  28. Sodek J, Ganss B, McKee MD. Osteopontin. Crit Rev Oral Biol Med. 2000;11:279–303.

    Article  CAS  Google Scholar 

  29. Darr H, Mayshar Y, Benvenisty N. Overexpression of NANOG in human ES cells enables feeder-free growth while inducing primitive ectoderm features. Development. 2006;133:1193–201.

    Article  CAS  Google Scholar 

  30. Armstrong L, Lako M, Lincoln J, Cairns PM, Hole N. mTert expression correlates with telomerase activity during the differentiation of murine embryonic stem cells. Mech Dev. 2000;97:109–16.

    Article  CAS  Google Scholar 

  31. Aubin JE. Regulation of osteoblast formation and function. Rev Endocr Metab Disord. 2001;2:81–94.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was afforded by the National Natural Science Foundation of China (No. 81571824), Natural Science Foundation of Jiangsu Province (BK20141141) and Fundamental Research Funds for the Central Universities (lzujbky-2018-27). The authors thank Dr. Ma Hongwei of the division of Nanobiomedicine, Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences for the instruments and Materials required for the QCM assay. The mESCs were kindly provided by Dr. Tang Fuchou at Peking University.

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Authors and Affiliations

  1. Laboratory of Biomaterials and Regenerative Medicine, Academy for Advanced Interdisciplinary Studies, Peking University, No. 5 Yi-He-Yuan Road, Beijing, 100871, People’s Republic of China

    Siqi Zhang, Yan Li, Zuyuan Luo, Ping Zhou & Shicheng Wei

  2. School and Hospital of Stomatology, Xuzhou Medical University, No. 209 Tong-Shan Road, Xuzhou, 221004, People’s Republic of China

    Yuhua Sun

  3. Central Laboratory, Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Peking University, No. 22 Zhong-Guan-Cun South Road, Hai-Dian District, Beijing, 100081, People’s Republic of China

    Yi Sui, Xiao Xu & Shicheng Wei

  4. School and Hospital of Stomatology, Lanzhou University, No. 222 Tian-Shui South Road, Lanzhou, 730000, People’s Republic of China

    Ping Zhou

  5. National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Peking University, No. 22 Zhong-Guan-Cun South Road, Hai-Dian District, Beijing, 100081, People’s Republic of China

    Shicheng Wei

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  1. Siqi Zhang
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Correspondence to Ping Zhou or Shicheng Wei.

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Zhang, S., Sun, Y., Sui, Y. et al. Determining Osteogenic Differentiation Efficacy of Pluripotent Stem Cells by Telomerase Activity. Tissue Eng Regen Med 15, 751–760 (2018). https://doi.org/10.1007/s13770-018-0138-6

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  • DOI: https://doi.org/10.1007/s13770-018-0138-6

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