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Efficient iPS Cell Generation from Blood Using Episomes and HDAC Inhibitors
Published on: October 28, 2014
DNp73 improves generation efficiency of human induced pluripotent stem cells
Yi Lin1, Zuxin Cheng, Zhijian Yang
1Stem Cell Research Center, Fujian Agriculture and Forestry University, Cangshan District, Fuzhou, Fujian, PR China.
Background:
Recent studies have found that p53 and its' associated cell cycle pathways are major inhibitors of human induced pluripotent stem (iPS) cell generation. In the same family as p53 is p73, which shares sequence similarities with p53. However, p73 also has distinct properties of its own, such as two alternative promoters to express transactivation of p73 (TAp73) and N terminal deleted p73 (DNp73). Functionally, TAp73 acts similarly to p53 in tumor suppression. However, DNp73, on the other hand acts as an oncogene to suppress p53 and p73 induced apoptosis. Therefore, how can p73 have opposing roles in human iPS cell generation?
Results:
Transcription factors, Oct4, Sox2, Klf4 and cMyc (4TF, Yamanaka factors) are used as basal conditions to generate iPS cells. In addition, the factor of DNp73(actually alpha splicing DNp73, DNp73α) is used to generate iPS cells. The experiment found that the addition of DNp73 gene increases human iPS cell generation efficiency by 12.6 folds in comparison to human fibroblast cells transduced with only the basal conditions. Also, iPS cells generated with DNp73 expression are more resistant to in vitro and in vivo differentiation.
Conclusions:
This study found DNp73, a family member of p53, is also involved in the human iPS cell generation. Specifically, that the involvement of DNp73 generates iPS cells that are more resistant to in vitro and in vivo differentiation. Therefore, this data may prove to be useful in future developmental studies and cancer researches.
Insights
The N-terminal deleted p73 (DNp73) gene significantly enhances human induced pluripotent stem (iPS) cell generation. These DNp73-generated iPS cells exhibit increased resistance to differentiation, offering potential for developmental and cancer research.
Area of Science:
- Stem Cell Biology
- Molecular Biology
- Cancer Research
Background:
- p53 and associated cell cycle pathways inhibit human induced pluripotent stem (iPS) cell generation.
- p73, a p53 family member, has distinct isoforms: TAp73 (tumor suppressor) and DNp73 (oncogene).
- DNp73's role in iPS cell generation, despite its oncogenic properties, remains unclear.
Purpose of the Study:
- To investigate the role of p73, specifically DNp73, in human iPS cell generation.
- To determine if DNp73 influences the efficiency and characteristics of generated iPS cells.
Main Methods:
- Human fibroblast cells were transduced with basal conditions (Oct4, Sox2, Klf4, cMyc - 4TF) for iPS cell generation.
- The DNp73 gene (specifically DNp73α) was added to the basal conditions.
- Comparison of iPS cell generation efficiency and differentiation resistance between groups.
Main Results:
- Addition of DNp73 increased human iPS cell generation efficiency by 12.6-fold compared to basal conditions alone.
- iPS cells generated with DNp73 expression showed enhanced resistance to both in vitro and in vivo differentiation.
- DNp73's involvement in iPS cell generation was demonstrated.
Conclusions:
- DNp73 plays a significant role in enhancing human iPS cell generation efficiency.
- DNp73-derived iPS cells possess superior resistance to differentiation.
- Findings suggest potential applications in future developmental biology and cancer research.
