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Bone morphogenetic protein 9 (BMP9) induces effective bone formation from reversibly immortalized multipotent adipose-derived (iMAD) mesenchymal stem cells.
Am J Transl Res 2016; 8(9):3710-3730AJ

Abstract

Regenerative medicine and bone tissue engineering using mesenchymal stem cells (MSCs) hold great promise as an effective approach to bone and skeletal reconstruction. While adipose tissue harbors MSC-like progenitors, or multipotent adipose-derived cells (MADs), it is important to identify and characterize potential biological factors that can effectively induce osteogenic differentiation of MADs. To overcome the time-consuming and technically challenging process of isolating and culturing primary MADs, here we establish and characterize the reversibly immortalized mouse multipotent adipose-derived cells (iMADs). The isolated mouse primary inguinal MAD cells are reversibly immortalized via the retrovirus-mediated expression of SV40 T antigen flanked with FRT sites. The iMADs are shown to express most common MSC markers. FLP-mediated removal of SV40 T antigen effectively reduces the proliferative activity and cell survival of iMADs, indicating the immortalization is reversible. Using the highly osteogenic BMP9, we find that the iMADs are highly responsive to BMP9 stimulation, express multiple lineage regulators, and undergo osteogenic differentiation in vitro upon BMP9 stimulation. Furthermore, we demonstrate that BMP9-stimulated iMADs form robust ectopic bone with a thermoresponsive biodegradable scaffold material. Collectively, our results demonstrate that the reversibly immortalized iMADs exhibit the characteristics of multipotent MSCs and are highly responsive to BMP9-induced osteogenic differentiation. Thus, the iMADs should provide a valuable resource for the study of MAD biology, which would ultimately enable us to develop novel and efficacious strategies for MAD-based bone tissue engineering.

Authors+Show Affiliations

Shandong Provincial Orthopaedics Hospital, The Provincial Hospital Affiliated to Shandong UniversityJinan 250021, China; Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical UniversityChongqing 400016, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Department of Biomedical Engineering, School of Bioengineering, Chongqing UniversityChongqing 400044, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical UniversityChongqing 400016, China.Department of Biomedical Engineering and Simpson Querrey Institute for BioNanotechnology in Medicine, Northwestern University Evanston, IL 60208, USA.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical UniversityChongqing 400016, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Departments of Surgery, Conservative Dentistry and Endodontics, West China Hospital and West China School of Stomatology, Sichuan UniversityChengdu 610041, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Department of General Surgery and Neurosurgery, The Affiliated Zhongnan Hospital of Wuhan UniversityWuhan 430071, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical UniversityChongqing 400016, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical UniversityChongqing 400016, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical UniversityChongqing 400016, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center Chicago, IL 60637, USA.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center Chicago, IL 60637, USA.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Departments of Surgery, Conservative Dentistry and Endodontics, West China Hospital and West China School of Stomatology, Sichuan UniversityChengdu 610041, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical UniversityChongqing 400016, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical UniversityChongqing 400016, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical UniversityChongqing 400016, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical UniversityChongqing 400016, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical UniversityChongqing 400016, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Department of Orthopaedic Surgery, Union Hospital of Tongji Medical College, Huazhong University of Science & TechnologyWuhan 430022, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Department of Biomedical Engineering, School of Bioengineering, Chongqing UniversityChongqing 400044, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Department of General Surgery and Neurosurgery, The Affiliated Zhongnan Hospital of Wuhan UniversityWuhan 430071, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical UniversityChongqing 400016, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center Chicago, IL 60637, USA.Section of Plastic Surgery, Department of Surgery, The University of Chicago Medical Center Chicago, IL 60637, USA.Department of Biomedical Engineering and Simpson Querrey Institute for BioNanotechnology in Medicine, Northwestern UniversityEvanston, IL 60208, USA; Department of Surgery, Feinberg School of MedicineChicago, IL 60616, USA.Shandong Provincial Orthopaedics Hospital, The Provincial Hospital Affiliated to Shandong University Jinan 250021, China.Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical CenterChicago, IL 60637, USA; Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical UniversityChongqing 400016, China.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

27725853

Citation

Lu, Shun, et al. "Bone Morphogenetic Protein 9 (BMP9) Induces Effective Bone Formation From Reversibly Immortalized Multipotent Adipose-derived (iMAD) Mesenchymal Stem Cells." American Journal of Translational Research, vol. 8, no. 9, 2016, pp. 3710-3730.
Lu S, Wang J, Ye J, et al. Bone morphogenetic protein 9 (BMP9) induces effective bone formation from reversibly immortalized multipotent adipose-derived (iMAD) mesenchymal stem cells. Am J Transl Res. 2016;8(9):3710-3730.
Lu, S., Wang, J., Ye, J., Zou, Y., Zhu, Y., Wei, Q., ... He, T. (2016). Bone morphogenetic protein 9 (BMP9) induces effective bone formation from reversibly immortalized multipotent adipose-derived (iMAD) mesenchymal stem cells. American Journal of Translational Research, 8(9), pp. 3710-3730.
Lu S, et al. Bone Morphogenetic Protein 9 (BMP9) Induces Effective Bone Formation From Reversibly Immortalized Multipotent Adipose-derived (iMAD) Mesenchymal Stem Cells. Am J Transl Res. 2016;8(9):3710-3730. PubMed PMID: 27725853.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Bone morphogenetic protein 9 (BMP9) induces effective bone formation from reversibly immortalized multipotent adipose-derived (iMAD) mesenchymal stem cells. AU - Lu,Shun, AU - Wang,Jing, AU - Ye,Jixing, AU - Zou,Yulong, AU - Zhu,Yunxiao, AU - Wei,Qiang, AU - Wang,Xin, AU - Tang,Shengli, AU - Liu,Hao, AU - Fan,Jiaming, AU - Zhang,Fugui, AU - Farina,Evan M, AU - Mohammed,Maryam M, AU - Song,Dongzhe, AU - Liao,Junyi, AU - Huang,Jiayi, AU - Guo,Dan, AU - Lu,Minpeng, AU - Liu,Feng, AU - Liu,Jianxiang, AU - Li,Li, AU - Ma,Chao, AU - Hu,Xue, AU - Lee,Michael J, AU - Reid,Russell R, AU - Ameer,Guillermo A, AU - Zhou,Dongsheng, AU - He,Tongchuan, Y1 - 2016/09/15/ PY - 2016/02/28/received PY - 2016/04/25/accepted PY - 2016/10/12/entrez PY - 2016/10/12/pubmed PY - 2016/10/12/medline KW - BMP9 KW - adipose-derived stem cells KW - bone formation KW - immortalized progenitor cells KW - mesenchymal stem cells KW - tissue engineering SP - 3710 EP - 3730 JF - American journal of translational research JO - Am J Transl Res VL - 8 IS - 9 N2 - Regenerative medicine and bone tissue engineering using mesenchymal stem cells (MSCs) hold great promise as an effective approach to bone and skeletal reconstruction. While adipose tissue harbors MSC-like progenitors, or multipotent adipose-derived cells (MADs), it is important to identify and characterize potential biological factors that can effectively induce osteogenic differentiation of MADs. To overcome the time-consuming and technically challenging process of isolating and culturing primary MADs, here we establish and characterize the reversibly immortalized mouse multipotent adipose-derived cells (iMADs). The isolated mouse primary inguinal MAD cells are reversibly immortalized via the retrovirus-mediated expression of SV40 T antigen flanked with FRT sites. The iMADs are shown to express most common MSC markers. FLP-mediated removal of SV40 T antigen effectively reduces the proliferative activity and cell survival of iMADs, indicating the immortalization is reversible. Using the highly osteogenic BMP9, we find that the iMADs are highly responsive to BMP9 stimulation, express multiple lineage regulators, and undergo osteogenic differentiation in vitro upon BMP9 stimulation. Furthermore, we demonstrate that BMP9-stimulated iMADs form robust ectopic bone with a thermoresponsive biodegradable scaffold material. Collectively, our results demonstrate that the reversibly immortalized iMADs exhibit the characteristics of multipotent MSCs and are highly responsive to BMP9-induced osteogenic differentiation. Thus, the iMADs should provide a valuable resource for the study of MAD biology, which would ultimately enable us to develop novel and efficacious strategies for MAD-based bone tissue engineering. SN - 1943-8141 UR - https://www.unboundmedicine.com/medline/citation/27725853/Bone_morphogenetic_protein_9__BMP9__induces_effective_bone_formation_from_reversibly_immortalized_multipotent_adipose_derived__iMAD__mesenchymal_stem_cells_ L2 - https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/27725853/ DB - PRIME DP - Unbound Medicine ER -