Figure S6. Downregulation of miR-10a or overexpression of KLF4 in old hBM-MSCs increased apoptotic gene expression. Quantification of mRNA expression of BAX and PUMA (pr-apoptotic), BCL2 and MCL1 (antiapoptotic) in O, O-anti10a and O-KLF4 hBM-MSCs after culture for 72 h under hypoxia conditions. n = 6/group. Mean ± SD. *P < 0.05 (PDF 38 kb

Chaowei Tian (5319839)

Chongyu Zhang (3192726)

Chuanfan Cheng (5319842)

Hongshen Huang (5319845)

Jianwei Liu (163716)

Jiao Li (261359)

Jun Dong (43792)

Pei Mo (5319836)

Weizhao Huang (5319848)

Zhenhui Zhang (847340)

Stem Cell Research & Therapy

Figure S3. Overexpression of miR-10a in old hBM-MSCs decreased apoptotic gene expression. Quantification of mRNA expression of BAX and PUMA (proapoptotic), BCL2 and MCL1 (antiapoptotic) in O and O-10a hBM-MSCs after culture for 72 h under hypoxia conditions. n = 6/group. Mean ± SD. *P < 0.05 (PDF 37 kb

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Additional file 4: of miR-10a rejuvenates aged human mesenchymal stem cells and improves heart function after myocardial infarction through KLF4

Figure S4. Overexpression of miR-10a in both young and old hBM-MSCs decreased hypoxia-induced apoptosis and increased cell survival. miR-10a transduced into young (Y-10a) and old (O-10a) hBM-MSCs by lentiviral vector. Control vector-transduced young hBM-MSCs (Y-c) and old hBM-MSCs (O-c) served as controls. Cells cultured for 72 h under hypoxia conditions. (A) Cell apoptosis assayed by TUNEL staining. Percentage of apoptotic cells (TUNEL+) quantified in Y-c, Y-10a, O-c, and O-10ahBM-MSCs. (B) Cell survival evaluated in Y-c, Y-10a, O-c, and O-10a hBM-MSCs. n = 6/group. Mean ± SD. *P < 0.05, Y-c vs Y-10a; #P < 0.05, O-c vs O-10a (PDF 94 kb

Additional file 5: of miR-10a rejuvenates aged human mesenchymal stem cells and improves heart function after myocardial infarction through KLF4

Figure S1. Proapoptotic and antiapoptotic gene expression in old hBM-MSCs under hypoxia conditions. Quantification of mRNA expression of BAX and PUMA (proapoptotic), BCL2 and MCL1 (antiapoptotic) in Y and O hBM-MSCs. n = 6/group. Mean ± SD. *P < 0.05 (PDF 36 kb

Additional file 2: of miR-10a rejuvenates aged human mesenchymal stem cells and improves heart function after myocardial infarction through KLF4

Figure S2. Expression of miR-10a and KLF4 in old hBM-MSCs regulated by lentiviral vector. Lentiviral vector carrying miR-10a sequence used to transduce old hBM-MSCs (O-10a) and control vector-transduced old hBM-MSCs (O-c) served as control. miR-10a expression was significantly higher in O-10a than in control vector-transduced (O) hBM-MSCs (A). Lentiviral vector carrying KLF4 siRNA sequence used to transduce old hBM-MSCs (O-anti-KLF4). KLF4 expression was significantly lower in O-antiKLF4 than in control vector-transduced O hBM-MSCs (B). Lentiviral vector carrying anti-miR-10a sequence used to transduce old hBM-MSCs (O-anti10a). miR-10a expression was significantly lower in O-anti10a than in control vector-transduced O hBM-MSCs (C). Lentiviral vector carrying KLF4 sequence used to transduce old hBM-MSCs (O-KLF4). KLF4 expression was significantly higher in O-KLF4 than in control vector-transduced O hBM-MSCs (D). Lentivirus which carries KLF4 vector used to infect miR-10a-upregulated old hBM-MSCs (O-10a) to restore KLF4 expression (O-10a-KLF4). miR-10a-upregulated old hBM-MSCs (O-10a) also infected by the control lentivirus (O-10a-c). KLF4 expression restored in O-10a-KLF4 compared to O-10a-c hBM-MSCs (E). n = 5/group. Mean ± SD. *P < 0.05 O-c vs O-10a, O-anti-KLF4, O-anti-10a, and O-KLF4; #P < 0.05 O-10a-KLF4 vs O-10a-c and O-10a (PDF 41 kb

Additional file 3: of miR-10a rejuvenates aged human mesenchymal stem cells and improves heart function after myocardial infarction through KLF4

Figure S8. miR-10a overexpression increased old hBM-MSC survival and decreased apoptosis by activating AKT. Expression of Akt phosphorylation in miR-10a-overexpressed old hBM-MSCs (O-10a) inhibited by Akt Inhibitor VI during 72-h culture under hypoxia conditions. (A) Expression of phosphor-(ser473)-AKT detected in O-c, O-10a, and Akt Inhibitor VI added O-10a hBM-MSCs (O-10a-P-AKT Inh). (B) Cell apoptosis assayed by TUNEL staining. Percentage of apoptotic cells (TUNEL+) quantified in O-c, O-10a, and O-10a-P-AKT Inh hBM-MSCs. (C) Cell survival evaluated in O-c, O-10a, and O-10a-P-AKT Inh hBM-MSCs. n = 5/group. Mean ± SD. *P < 0.05, O-10a-P-AKT Inh vs O-10a (PDF 184 kb

Additional file 9: of miR-10a rejuvenates aged human mesenchymal stem cells and improves heart function after myocardial infarction through KLF4

Additional file 7: of miR-10a rejuvenates aged human mesenchymal stem cells and improves heart function after myocardial infarction through KLF4

Table S1. qRT-PCR primer and miRNA RT primer sequences (DOCX 19Â kb

Additional file 1: of miR-10a rejuvenates aged human mesenchymal stem cells and improves heart function after myocardial infarction through KLF4

Figure S9. Expression of VEGF and SDF mRNA in mouse hearts after MI and hBM-MSCs. (A) Expression of VEGF and SDF mRNA determined in border region of infarcted mouse hearts that received implantation of control vector-transduced young hBM-MSCs (Y-c), control vector-transduced old hBM-MSCs (O-c), miR-10a-overexpressed old hBM-MSCs (O-10a), or KLF4-inhibited old hBM-MSCs (O-antiKLF4) following MI. (B) Expression of VEGF and SDF mRNA determined by RT-qPCR in Y-c, O-c, O-10a, or O-antiKLF4 hBM-MSCs after hypoxia for 72 h. n = 6/group. Mean ± SD. *P < 0.05, Y-c vs other groups; #P < 0.05, O-c vs other groups (PDF 39 kb

Additional file 10: of miR-10a rejuvenates aged human mesenchymal stem cells and improves heart function after myocardial infarction through KLF4

Figure S7. Antiapoptotic effect of miR-10a attenuated by restoration of KLF4 quantified by RT-qPCR. Quantification of mRNA expression of BAX and PUMA (proapoptotic), BCL2 and MCL1 (antiapoptotic) in O-10a-c, O-10a-KLF4, and O-c hBM-MSCs after culture for 72 h under hypoxia conditions. n = 6/group. Mean ± SD. *P < 0.05 (PDF 57 kb

Additional file 8: of miR-10a rejuvenates aged human mesenchymal stem cells and improves heart function after myocardial infarction through KLF4

Abstract Background Aging is one of the key factors that regulate the function of human bone marrow mesenchymal stem cells (hBM-MSCs) and related changes in microRNA (miRNA) expression. However, data reported on aging-related miRNA changes in hBM-MSCs are limited. Methods We demonstrated previously that miR-10a is significantly decreased in aged hBM-MSCs and restoration of the miR-10a level attenuated cell senescence and increased the differentiation capacity of aged hBM-MSCs by repressing Krüpple-like factor 4 (KLF4). In the present study, miR-10a was overexpressed or KLF4 was downregulated in old hBM-MSCs by lentiviral transduction. The hypoxia-induced apoptosis, cell survival, and cell paracrine function of aged hBM-MSCs were investigated in vitro. In vivo, miR-10a-overexpressed or KLF4-downregulated old hBM-MSCs were implanted into infarcted mouse hearts after myocardial infarction (MI). The mouse cardiac function of cardiac angiogenesis was measured and cell survival of aged hBM-MSCs was investigated. Results Through lentivirus-mediated upregulation of miR-10a and downregulation of KLF4 in aged hBM-MSCs in vitro, we revealed that miR-10a decreased hypoxia-induced cell apoptosis and increased cell survival of aged hBM-MSCs by repressing the KLF4–BAX/BCL2 pathway. In vivo, transplantation of miR-10a-overexpressed aged hBM-MSCs promoted implanted stem cell survival and improved cardiac function after MI. Mechanistic studies revealed that overexpression of miR-10a in aged hBM-MSCs activated Akt and stimulated the expression of angiogenic factors, thus increasing angiogenesis in ischemic mouse hearts. Conclusions miR-10a rejuvenated aged hBM-MSCs which improved angiogenesis and cardiac function in injured mouse hearts

Jun Dong

Zhenhui Zhang

Hongshen Huang

Pei Mo

Chuanfan Cheng

Jianwei Liu

Weizhao Huang

Chaowei Tian

Chongyu Zhang

Jiao Li

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miR-10a rejuvenates aged human mesenchymal stem cells and improves heart function after myocardial infarction through KLF4

https://figshare.com/articles/Additional_file_8_of_miR-10a_rejuvenates_aged_human_mesenchymal_stem_cells_and_improves_heart_function_after_myocardial_infarction_through_KLF4/6395609

Additional file 7: of miR-10a rejuvenates aged human mesenchymal stem cells and improves heart function after myocardial infarction through KLF4

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