Current Issue

Volume 21, Number 2, Jul-Sep (Summer) 2019 , Serial Number: 82 Pages: 161-168

Generation and Characterization of Induced Pluripotent Stem Cells from Mononuclear Cells in Schizophrenic Patients


Qing Liu, M.D, 1, 2, 3, #, Jiang Du, Ph.D, 1, 4, #, Jinyu Fan, Ph.D, 1, 2, Wenqiang Li, Ph.D, 1, 3, Weiyun Guo, M.D, 2, Huigen Feng, Ph.D., 1, 2, *, Juntang Lin, Ph.D., 1, 4, *,
Stem Cell and Biotherapy Engineering Research Center of Henan, Xinxiang Medical University, Xinxiang, China
College of Life Science and Technology, Xinxiang Medical University, Xinxiang, China
Henan Key Lab of Biological Psychiatry, The Second Affiliated Hospital of Xinxiang Medical University, Xinxiang, China
College of Biomedical Engineering, Xinxiang Medical University, Xinxiang, China
*Corresponding Address: Stem Cell and Biotherapy Engineering Research Center of Henan Xinxiang Medical University Xinxiang China Emails:fenghuigen@xxmu.edu.cn,linjtlin@126.com

The first two authors contributed equally to this work.

Abstract

Objective

Schizophrenia (SZ) is a mental disorder in which psychotic symptoms are the main problem. The pathogenesis of SZ is not fully understood, partly because of limitations in current disease models and technology. The development of induced pluripotent stem cell (iPSC) technology has opened up the possibility of elucidating disease mechanisms in neurodegenerative diseases. Here, we aimed to obtain iPSCs from peripheral blood mononuclear cells (PBMCs) of normal and schizophrenic individuals and analyze the inflammatory response in these iPSCs.

Materials and Methods

In this experimental study, we isolated PBMCs from whole blood of healthy individuals and SZ patients and reprogrammed them into iPSCs by transfection of recombinant lentiviruses that contained Yamanaka factors (Oct4, Sox2, Klf4 and c-Myc). We calculated the numbers of iPSC clones and stained them with alkaline phosphatase (ALP), Nanog, SSEA4, Nestin, Vimentin, and AFP to confirm their efficiency and pluripotency. The iPSCs were analyzed by real-time quantitative polymerase chain reaction (qRT-PCR) for the expressions of inflammatory factors.

Results

iPSCs from schizophrenic patients (SZ-iPSCs) exhibited typical morphology and highly expressed pluripotent markers. These iPSCs retained their normal karyotype and differentiated in vitro to form embryoid bodies (EBs) that expressed markers of all 3 germ layers. However, iPSCs from the SZ-iPSCs group had a weak capacity to differentiate into ectoderm compared to the normal iPSCs (Con-iPSC). An elevated, stronger inflammatory response existed in iPSCs from schizophrenic individuals.

Conclusion

We successfully obtained iPSCs from PBMCs of schizophrenic patients without genetic operation and analyzed the expressions of pluripotent markers and inflammatory factors between the Con-iPSC and SZ-iPSC groups. Taken together, our results may assist to explain the pathogenesis of SZ and develop new strategies for clinical diagnosis and treatment.