Transcriptional profiling of human mesenchymal stem cells transduced with reporter genes for imaging

Physiological Genomics - Tập 37 Số 1 - Trang 23-34 - 2009
Fangjing Wang1, James E. Dennis2, Amad Awadallah2, Luis A. Solchaga3,4, Joseph Molter5, Yu Kuang1, Nicolas Salem1, Yuan Lin4, Haibin Tian5, Jeffery A. Kolthammer1, Yunhui Kim5, Zachary Love5, Stanton L. Gerson3,4, Zhenghong Lee3,1,5
1Department of Biomedical Engineering, University Hospitals, Case Medical Center, Case Western Reserve University, Cleveland, Ohio
2Department of Orthopedics, University Hospitals, Case Medical Center, Case Western Reserve University, Cleveland, Ohio
3Center for Stem Cell and Regenerative Medicine, University Hospitals, Case Medical Center, Case Western Reserve University, Cleveland, Ohio
4Department of Hematology/Oncology, University Hospitals, Case Medical Center, Case Western Reserve University, Cleveland, Ohio
5Department of Nuclear Medicine/Radiology, University Hospitals, Case Medical Center, Case Western Reserve University, Cleveland, Ohio

Tóm tắt

Mesenchymal stem cells (MSCs) can differentiate into osteogenic, adipogenic, chondrogenic, myocardial, or neural lineages when exposed to specific stimuli, making them attractive for tissue repair and regeneration. We have used reporter gene-based imaging technology to track MSC transplantation or implantation in vivo. However, the effects of lentiviral transduction with the fluc-mrfp-ttk triple-fusion vector on the transcriptional profiles of MSCs remain unknown. In this study, gene expression differences between wild-type and transduced hMSCs were evaluated using an oligonucleotide human microarray. Significance Analysis of Microarray identified differential genes with high accuracy; RT-PCR validated the microarray results. Annotation analysis showed that transduced hMSCs upregulated cell differentiation and antiapoptosis genes while downregulating cell cycle, proliferation genes. Despite transcriptional changes associated with bone and cartilage remodeling, their random pattern indicates no systematic change of crucial genes that are associated with osteogenic, adipogenic, or chondrogenic differentiation. This correlates with the experimental results that lentiviral transduction did not cause the transduced MSCs to lose their basic stem cell identity as demonstrated by osteogenic, chondrogenic, and adipogenic differentiation assays with both transduced and wild-type MSCs, although a certain degree of alterations occurred. Histological analysis demonstrated osteogenic differentiation in MSC-loaded ceramic cubes in vivo. In conclusion, transduction of reporter genes into MSCs preserved the basic properties of stem cells while enabling noninvasive imaging in living animals to study the biodistribution and other biological activities of the cells.

Từ khóa


Tài liệu tham khảo

10.1182/blood-2004-02-0655

10.1006/excr.2001.5278

10.1007/s00259-006-0093-7

10.1002/(SICI)1097-4644(199702)64:2<278::AID-JCB11>3.0.CO;2-F

10.1016/S0076-6879(04)86013-0

10.1161/CIRCULATIONAHA.105.588954

10.1002/jor.1100090504

Carter D, Tyrell A, Bubnic S, Marcelino M, Kedzierski K, Monroy R, Mills R, Danilkovitch A.Characterization of MSC potential to treat GVHD using molecular markers linked to MSC-mediated immunosuppression in vitro.Blood106: 160B–160B, 2005.

Daley GQ, Goodell MA, Snyder EY.Realistic prospects for stem cell therapeutics.Hematology Am Soc Hematol Educ Program398–418, 2003.

10.1182/blood-2002-06-1830

10.1006/bbrc.2001.5543

10.1159/000047856

10.1002/jgm.729

10.1161/01.CIR.0000163546.27639.AA

10.1038/6529

10.1016/j.gene.2004.06.028

10.1074/jbc.275.13.9645

10.1038/nature00870

10.1088/0031-9155/50/19/001

Johnstone B, Yoo JU.Autologous mesenchymal progenitor cells in articular cartilage repair.Clin Orthop Relat ResS156–S162, 1999.

Jundt G, Berghauser KH, Termine JD, Schulz A.Osteonectin–a differentiation marker of bone cells.Cell Tissue Res248: 409–415, 1987.

10.1074/jbc.M413834200

Keilhoff G, Stang F, Goihl A, Wolf G, Fansa H.Transdifferentiated mesenchymal stem cells as alternative therapy in supporting nerve regeneration and myelination.Cell Mol Neurobiol26: 1235–1252, 2006.

10.1200/JCO.2000.18.2.307

10.1096/fj.05-4551fje

10.1212/WNL.59.4.514

10.2967/jnumed.107.043166

10.1016/j.nucmedbio.2005.04.018

10.1242/jcs.02932

10.1073/pnas.0508945102

10.1158/0008-5472.CAN-04-1874

10.1532/IJH97.07041

10.1002/jcp.1041430304

10.1007/s00259-003-1441-5

10.1111/j.1440-1797.2006.00660.x

10.1073/pnas.0532693100

10.1038/nm0603-702

10.1158/0008-5472.CAN-03-1816

Sarrazin P, Hackett JA, Fortier I, Gallant MA, de Brum-Fernandes A.Role of EP3 and EP4 prostaglandin receptors in reorganization of the cytoskeleton in mature human osteoclasts.J Rheumatol31: 1598–1606, 2004.

10.1074/jbc.M412630200

10.1291/hypres.24.437

10.1016/S0049-3848(98)00150-9

Wang FJ, Anderson PW, Salem N, Kuang Y, Tennant BC, Lee Z.Gene expression studies of hepatitis virus-induced woodchuck hepatocellular carcinoma in correlation with human results.Int J Oncol30: 33–44, 2007.

10.1152/physiolgenomics.00254.2005

10.1002/jor.1100160403

10.1002/jcb.10669

10.1006/mthe.2002.0571

Thông tin
Thông tin xuất bản

Physiological Genomics

Tập 37 Số 1

23-34

Thông tin tác giả