PMID- 30850008 OWN - NLM STAT- MEDLINE DCOM- 20200422 LR - 20200422 IS - 1757-6512 (Electronic) IS - 1757-6512 (Linking) VI - 10 IP - 1 DP - 2019 Mar 8 TI - In vitro expansion affects the response of human bone marrow stromal cells to irradiation. PG - 82 LID - 10.1186/s13287-019-1191-3 [doi] LID - 82 AB - BACKGROUND: Bone marrow stromal cells (BMSCs) are extensively used in regeneration therapy and cytology experiments simulate how BMSCs respond to radiation. Due to the small number and the heterogeneity of primary isolated BMSCs, extensive in vitro expansion is usually required before application, which affects the cellular characteristics and gene expression of BMSCs. However, whether the radiation response of BMSCs changes during in vitro expansion is unclear. METHODS: In this study, BMSCs were passaged in vitro and irradiated at passage 6 (P6) and passage 10 (P10). Then, apoptosis, the cell cycle, senescence, the cytokine secretion and the gene expression profile were analysed for the P6, P10, and non-irradiated (control) BMSCs at different post-irradiation time points. RESULTS: The P6 BMSCs had a lower percentage of apoptotic cells than the P10 BMSCs at 24 and 48 h post-irradiation but not compared to that of the controls at 2 and 8 h post-irradiation. The P6 BMSCs had a lower percentage of cells in S phase and a higher percentage in G1 phase than the P10 BMSCs at 2 and 8 h post-irradiation. The radiation had similar effects on the senescent cell level and impaired immunomodulation capacity of the P6 and P10 BMSCs. Regardless of whether they were irradiated, the P6 and P10 BMSCs always expressed a distinctive set of genes. The upregulated genes were enriched in pathways including the cell cycle, DNA replication and oocyte meiosis. Then, a subset of conserved irradiation response genes across the BMSCs was identified, comprising 12 differentially upregulated genes and 5 differentially downregulated genes. These genes were especially associated with the p53 signaling pathway, DNA damage and DNA repair. Furthermore, validation experiments revealed that the mRNA and protein levels of these conserved genes were different between the P6 and P10 BMSCs after irradiation. Weighted gene co-expression network analysis supported these findings and further revealed the effects of cell passage on the irradiation response in BMSCs. CONCLUSION: The results indicated that cell passage in vitro affected the irradiation response of BMSCs via molecular mechanisms that mediated differences in apoptosis, the cell cycle, senescence and the cytokine secretion. Thus, accurate cell passage information is not only important for transplantation therapy but also for future studies on the radiation response in BMSCs. FAU - Xiang, Yang AU - Xiang Y AD - Department of Blood Transfusion, Irradiation biology laboratory, The Second Affiliated Hospital, Army Medical University, Xinqiao Road, Shapingba, Chongqing, 400037, China. FAU - Wu, Chun AU - Wu C AD - Department of Blood Transfusion, Irradiation biology laboratory, The Second Affiliated Hospital, Army Medical University, Xinqiao Road, Shapingba, Chongqing, 400037, China. AD - Central Laboratory, The Second Affiliated Hospital, Army Medical University, Xinqiao Road, Shapingba, Chongqing, 400037, China. FAU - Wu, Jiang AU - Wu J AD - Department of Blood Transfusion, Irradiation biology laboratory, The Second Affiliated Hospital, Army Medical University, Xinqiao Road, Shapingba, Chongqing, 400037, China. FAU - Quan, Weili AU - Quan W AD - Center for Genome Analysis, ABLife Inc., Wuhan, 430075, Hubei, China. FAU - Cheng, Chao AU - Cheng C AD - Center for Genome Analysis, ABLife Inc., Wuhan, 430075, Hubei, China. FAU - Zhou, Jian AU - Zhou J AD - Center for Genome Analysis, ABLife Inc., Wuhan, 430075, Hubei, China. FAU - Chen, Li AU - Chen L AD - Department of Blood Transfusion, Irradiation biology laboratory, The Second Affiliated Hospital, Army Medical University, Xinqiao Road, Shapingba, Chongqing, 400037, China. FAU - Xiang, Lixin AU - Xiang L AD - Department of Blood Transfusion, Irradiation biology laboratory, The Second Affiliated Hospital, Army Medical University, Xinqiao Road, Shapingba, Chongqing, 400037, China. FAU - Li, Fengjie AU - Li F AD - Department of Blood Transfusion, Irradiation biology laboratory, The Second Affiliated Hospital, Army Medical University, Xinqiao Road, Shapingba, Chongqing, 400037, China. FAU - Zhang, Kebin AU - Zhang K AD - Central Laboratory, The Second Affiliated Hospital, Army Medical University, Xinqiao Road, Shapingba, Chongqing, 400037, China. FAU - Ran, Qian AU - Ran Q AD - Department of Blood Transfusion, Irradiation biology laboratory, The Second Affiliated Hospital, Army Medical University, Xinqiao Road, Shapingba, Chongqing, 400037, China. louise-r-q@163.com. FAU - Zhang, Yi AU - Zhang Y AD - Center for Genome Analysis, ABLife Inc., Wuhan, 430075, Hubei, China. FAU - Li, Zhongjun AU - Li Z AD - Department of Blood Transfusion, Irradiation biology laboratory, The Second Affiliated Hospital, Army Medical University, Xinqiao Road, Shapingba, Chongqing, 400037, China. johnneyusc@gmail.com. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20190308 PL - England TA - Stem Cell Res Ther JT - Stem cell research & therapy JID - 101527581 SB - IM MH - Apoptosis/*radiation effects MH - Bone Marrow Cells/*metabolism MH - Cellular Senescence/*radiation effects MH - *Gamma Rays MH - Gene Expression Profiling MH - Gene Expression Regulation/*radiation effects MH - Humans MH - Stromal Cells/metabolism PMC - PMC6408817 OTO - NOTNLM OT - BMSCs OT - Cell passage OT - Irradiation OT - Transcriptome COIS- ETHICS APPROVAL AND CONSENT TO PARTICIPATE: The study was approved by the Army Medical University Institutional Ethics Committee. CONSENT FOR PUBLICATION: Not applicable. COMPETING INTERESTS: The authors declare that they have no competing interests. PUBLISHER'S NOTE: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. EDAT- 2019/03/10 06:00 MHDA- 2020/04/23 06:00 PMCR- 2019/03/08 CRDT- 2019/03/10 06:00 PHST- 2018/09/30 00:00 [received] PHST- 2019/02/25 00:00 [accepted] PHST- 2019/01/25 00:00 [revised] PHST- 2019/03/10 06:00 [entrez] PHST- 2019/03/10 06:00 [pubmed] PHST- 2020/04/23 06:00 [medline] PHST- 2019/03/08 00:00 [pmc-release] AID - 10.1186/s13287-019-1191-3 [pii] AID - 1191 [pii] AID - 10.1186/s13287-019-1191-3 [doi] PST - epublish SO - Stem Cell Res Ther. 2019 Mar 8;10(1):82. doi: 10.1186/s13287-019-1191-3.