Tissue Eng Regen Med.  2018 Dec;15(6):751-760. 10.1007/s13770-018-0138-6.

Determining Osteogenic Differentiation Efficacy of Pluripotent Stem Cells by Telomerase Activity

Affiliations
  • 1Laboratory 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. zhoup@lzu.edu.cn, sc-wei@pku.edu.cn
  • 2School and Hospital of Stomatology, Xuzhou Medical University, No. 209 Tong-Shan Road, Xuzhou 221004, People's Republic of China.
  • 3Central 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.
  • 4School and Hospital of Stomatology, Lanzhou University, No. 222 Tian-Shui South Road, Lanzhou 730000, People's Republic of China.
  • 5National 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.

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.

Keyword

Mouse embryonic stem cells; Pluripotency; Telomerase activity; Osteogenic differentiation; Efficiency

MeSH Terms

Bone and Bones
Down-Regulation
Gene Expression
In Vitro Techniques
Methods
Mouse Embryonic Stem Cells
Osteoblasts
Pluripotent Stem Cells*
Quartz Crystal Microbalance Techniques
Telomerase*
Up-Regulation
Telomerase
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