Format

Send to

Choose Destination
Stem Cells Int. 2017;2017:3167985. doi: 10.1155/2017/3167985. Epub 2017 Nov 5.

The Impact of GFP Reporter Gene Transduction and Expression on Metabolomics of Placental Mesenchymal Stem Cells Determined by UHPLC-Q/TOF-MS.

Yang J1,2, Wang N2,3, Chen D1,2, Yu J1,2, Pan Q1,2, Wang D1,2, Liu J1,2, Shi X4, Dong X1,2, Cao H1,2, Li L2,3, Li L1,2.

Author information

1
State Key Laboratory for the Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China.
2
Collaborative Innovation Center for the Diagnosis and Treatment of Infectious Diseases, Zhejiang University, Hangzhou 310003, China.
3
Department of Chemistry, University of Alberta, Edmonton, AB, Canada T6G 2G2.
4
Chu Kochen Honors College, Zhejiang University, 866 Yuhangtang Rd., Hangzhou 310058, China.

Abstract

Introduction:

Green fluorescent protein (GFP) is widely used as a reporter gene in regenerative medicine research to label and track stem cells. Here, we examined whether expressing GFP gene may impact the metabolism of human placental mesenchymal stem cells (hPMSCs).

Methods:

The GFP gene was transduced into hPMSCs using lentiviral-based infection to establish GFP+hPMSCs. A sensitive 13C/12C-dansyl labeling LC-MS method targeting the amine/phenol submetabolome was used for in-depth cell metabolome profiling.

Results:

A total of 1151 peak pairs or metabolites were detected from 12 LC-MS runs. Principal component analysis and partial least squares discriminant analysis showed poor separation, and the volcano plots demonstrated that most of the metabolites were not significantly changed when hPMSCs were tagged with GFP. Overall, 739 metabolites were positively or putatively identified. Only 11 metabolites showed significant changes. Metabolic pathway analyses indicated that three of the identified metabolites were involved in nine pathways. However, these metabolites are unlikely to have a large impact on the metabolic pathways due to their nonessential roles and limited hits in pathway analysis.

Conclusion:

This study indicated that the expression of ectopic GFP reporter gene did not significantly alter the metabolomics pathways covered by the amine/phenol submetabolome.

Supplemental Content

Full text links

Icon for Hindawi Limited Icon for PubMed Central
Loading ...
Support Center