Donor variation and loss of multipotency during in vitro expansion of human mesenchymal stem cells for bone tissue engineering
The use of multipotent human mesenchymal stem cells (hMSCs) for tissue engineering has been a subject of extensive research. The donor variation in growth, differentiation and in vivo bone forming ability of hMSCs is a bottleneck for standardization of therapeutic protocols. In this study, we isolat...
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| Published in | Journal of orthopaedic research Vol. 25; no. 8; pp. 1029 - 1041 |
|---|---|
| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
| Published |
Hoboken
Wiley Subscription Services, Inc., A Wiley Company
01.08.2007
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| Subjects | |
| Online Access | Get full text |
| ISSN | 0736-0266 1554-527X |
| DOI | 10.1002/jor.20402 |
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| Abstract | The use of multipotent human mesenchymal stem cells (hMSCs) for tissue engineering has been a subject of extensive research. The donor variation in growth, differentiation and in vivo bone forming ability of hMSCs is a bottleneck for standardization of therapeutic protocols. In this study, we isolated and characterized hMSCs from 19 independent donors, aged between 27 and 85 years, and investigated the extent of heterogeneity of the cells and the extent to which hMSCs can be expanded without loosing multipotency. Dexamethasone‐induced ALP expression varied between 1.2‐ and 3.7‐fold, but no correlation was found with age, gender, or source of isolation. The cells from donors with a higher percentage of ALP‐positive cells in control and dexamethasone‐induced groups showed more calcium deposition than cells with lower percentage of ALP positive cells. Despite the variability in osteogenic gene expression among the donors tested, ALP, Collagen type 1, osteocalcin, and S100A4 showed similar trends during the course of osteogenic differentiation. In vitro expansion studies showed that hMSCs can be effectively expanded up to four passages (approximately 10–12 population doublings from a P0 culture) while retaining their multipotency. Our in vivo studies suggest a correlation between in vitro ALP expression and in vivo bone formation. In conclusion, irrespective of age, gender, and source of isolation, cells from all donors showed osteogenic potential. The variability in ALP expression appears to be a result of sampling method and cellular heterogeneity among the donor population. © 2007 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 25:1029–1041, 2007 |
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| AbstractList | The use of multipotent human mesenchymal stem cells (hMSCs) for tissue engineering has been a subject of extensive research. The donor variation in growth, differentiation and in vivo bone forming ability of hMSCs is a bottleneck for standardization of therapeutic protocols. In this study, we isolated and characterized hMSCs from 19 independent donors, aged between 27 and 85 years, and investigated the extent of heterogeneity of the cells and the extent to which hMSCs can be expanded without loosing multipotency. Dexamethasone-induced ALP expression varied between 1.2- and 3.7-fold, but no correlation was found with age, gender, or source of isolation. The cells from donors with a higher percentage of ALP-positive cells in control and dexamethasone-induced groups showed more calcium deposition than cells with lower percentage of ALP positive cells. Despite the variability in osteogenic gene expression among the donors tested, ALP, Collagen type 1, osteocalcin, and S100A4 showed similar trends during the course of osteogenic differentiation. In vitro expansion studies showed that hMSCs can be effectively expanded up to four passages (approximately 10-12 population doublings from a P0 culture) while retaining their multipotency. Our in vivo studies suggest a correlation between in vitro ALP expression and in vivo bone formation. In conclusion, irrespective of age, gender, and source of isolation, cells from all donors showed osteogenic potential. The variability in ALP expression appears to be a result of sampling method and cellular heterogeneity among the donor population.The use of multipotent human mesenchymal stem cells (hMSCs) for tissue engineering has been a subject of extensive research. The donor variation in growth, differentiation and in vivo bone forming ability of hMSCs is a bottleneck for standardization of therapeutic protocols. In this study, we isolated and characterized hMSCs from 19 independent donors, aged between 27 and 85 years, and investigated the extent of heterogeneity of the cells and the extent to which hMSCs can be expanded without loosing multipotency. Dexamethasone-induced ALP expression varied between 1.2- and 3.7-fold, but no correlation was found with age, gender, or source of isolation. The cells from donors with a higher percentage of ALP-positive cells in control and dexamethasone-induced groups showed more calcium deposition than cells with lower percentage of ALP positive cells. Despite the variability in osteogenic gene expression among the donors tested, ALP, Collagen type 1, osteocalcin, and S100A4 showed similar trends during the course of osteogenic differentiation. In vitro expansion studies showed that hMSCs can be effectively expanded up to four passages (approximately 10-12 population doublings from a P0 culture) while retaining their multipotency. Our in vivo studies suggest a correlation between in vitro ALP expression and in vivo bone formation. In conclusion, irrespective of age, gender, and source of isolation, cells from all donors showed osteogenic potential. The variability in ALP expression appears to be a result of sampling method and cellular heterogeneity among the donor population. The use of multipotent human mesenchymal stem cells (hMSCs) for tissue engineering has been a subject of extensive research. The donor variation in growth, differentiation and in vivo bone forming ability of hMSCs is a bottleneck for standardization of therapeutic protocols. In this study, we isolated and characterized hMSCs from 19 independent donors, aged between 27 and 85 years, and investigated the extent of heterogeneity of the cells and the extent to which hMSCs can be expanded without loosing multipotency. Dexamethasone‐induced ALP expression varied between 1.2‐ and 3.7‐fold, but no correlation was found with age, gender, or source of isolation. The cells from donors with a higher percentage of ALP‐positive cells in control and dexamethasone‐induced groups showed more calcium deposition than cells with lower percentage of ALP positive cells. Despite the variability in osteogenic gene expression among the donors tested, ALP, Collagen type 1, osteocalcin, and S100A4 showed similar trends during the course of osteogenic differentiation. In vitro expansion studies showed that hMSCs can be effectively expanded up to four passages (approximately 10–12 population doublings from a P0 culture) while retaining their multipotency. Our in vivo studies suggest a correlation between in vitro ALP expression and in vivo bone formation. In conclusion, irrespective of age, gender, and source of isolation, cells from all donors showed osteogenic potential. The variability in ALP expression appears to be a result of sampling method and cellular heterogeneity among the donor population. © 2007 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 25:1029–1041, 2007 The use of multipotent human mesenchymal stem cells (hMSCs) for tissue engineering has been a subject of extensive research. The donor variation in growth, differentiation and in vivo bone forming ability of hMSCs is a bottleneck for standardization of therapeutic protocols. In this study, we isolated and characterized hMSCs from 19 independent donors, aged between 27 and 85 years, and investigated the extent of heterogeneity of the cells and the extent to which hMSCs can be expanded without loosing multipotency. Dexamethasone‐induced ALP expression varied between 1.2‐ and 3.7‐fold, but no correlation was found with age, gender, or source of isolation. The cells from donors with a higher percentage of ALP‐positive cells in control and dexamethasone‐induced groups showed more calcium deposition than cells with lower percentage of ALP positive cells. Despite the variability in osteogenic gene expression among the donors tested, ALP , Collagen type 1, osteocalcin , and S100A4 showed similar trends during the course of osteogenic differentiation. In vitro expansion studies showed that hMSCs can be effectively expanded up to four passages (approximately 10–12 population doublings from a P0 culture) while retaining their multipotency. Our in vivo studies suggest a correlation between in vitro ALP expression and in vivo bone formation. In conclusion, irrespective of age, gender, and source of isolation, cells from all donors showed osteogenic potential. The variability in ALP expression appears to be a result of sampling method and cellular heterogeneity among the donor population. © 2007 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 25:1029–1041, 2007 The use of multipotent human mesenchymal stem cells (hMSCs) for tissue engineering has been a subject of extensive research. The donor variation in growth, differentiation and in vivo bone forming ability of hMSCs is a bottleneck for standardization of therapeutic protocols. In this study, we isolated and characterized hMSCs from 19 independent donors, aged between 27 and 85 years, and investigated the extent of heterogeneity of the cells and the extent to which hMSCs can be expanded without loosing multipotency. Dexamethasone-induced ALP expression varied between 1.2- and 3.7-fold, but no correlation was found with age, gender, or source of isolation. The cells from donors with a higher percentage of ALP-positive cells in control and dexamethasone-induced groups showed more calcium deposition than cells with lower percentage of ALP positive cells. Despite the variability in osteogenic gene expression among the donors tested, ALP, Collagen type 1, osteocalcin, and S100A4 showed similar trends during the course of osteogenic differentiation. In vitro expansion studies showed that hMSCs can be effectively expanded up to four passages (approximately 10-12 population doublings from a P0 culture) while retaining their multipotency. Our in vivo studies suggest a correlation between in vitro ALP expression and in vivo bone formation. In conclusion, irrespective of age, gender, and source of isolation, cells from all donors showed osteogenic potential. The variability in ALP expression appears to be a result of sampling method and cellular heterogeneity among the donor population. |
| Author | Siddappa, Ramakrishnaiah de Boer, Jan Licht, Ruud van Blitterswijk, Clemens |
| Author_xml | – sequence: 1 givenname: Ramakrishnaiah surname: Siddappa fullname: Siddappa, Ramakrishnaiah organization: Institute for BioMedical Technology, Department of Tissue Regeneration, University of Twente, Zuidhorst, P.O. Box 217, Enschede 7500 AE, The Netherlands – sequence: 2 givenname: Ruud surname: Licht fullname: Licht, Ruud organization: Institute for BioMedical Technology, Department of Tissue Regeneration, University of Twente, Zuidhorst, P.O. Box 217, Enschede 7500 AE, The Netherlands – sequence: 3 givenname: Clemens surname: van Blitterswijk fullname: van Blitterswijk, Clemens organization: Institute for BioMedical Technology, Department of Tissue Regeneration, University of Twente, Zuidhorst, P.O. Box 217, Enschede 7500 AE, The Netherlands – sequence: 4 givenname: Jan surname: de Boer fullname: de Boer, Jan email: j.deboer@tnw.utwente.nl organization: Institute for BioMedical Technology, Department of Tissue Regeneration, University of Twente, Zuidhorst, P.O. Box 217, Enschede 7500 AE, The Netherlands |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/17469183$$D View this record in MEDLINE/PubMed |
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| References | Pola E, Gao W, Zhou Y, et al. 2004. Efficient bone formation by gene transfer of human LIM mineralization protein-3. Gene Ther 11: 683-693. Bancroft GN, Sikavitsas VI, van den Dolder J, et al. 2002. Fluid flow increases mineralized matrix deposition in 3D perfusion culture of marrow stromal osteoblasts in a dose-dependent manner. Proc Natl Acad Sci USA 99: 12600-12605. Bruder SP, Jaiswal N, Haynesworth SE. 1997. Growth kinetics, self-renewal, and the osteogenic potential of purified human mesenchymal stem cells during extensive subcultivation and following cryopreservation. J Cell Biochem 64: 278-294. Fehrer C, Lepperdinger G. 2005. Mesenchymal stem cell aging. Exp Gerontol 12: 926-930. Cowan CM, et al. 2004. Adipose-derived adult stromal cells heal critical-size mouse calvarial defects. Nat Biotechnol 22: 560-567. D'Ippolito G, Schiller PC, Ricordi C, et al. 1999. Age-related osteogenic potential of mesenchymal stromal stem cells from human vertebral bone marrow. J Bone Miner Res 14: 1115-1122. Livak KJ, Schmittgen TD. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25: 402-408. Le Blanc K, Pittenger M. 2005. Mesenchymal stem cells: progress toward promise. Cytotherapy 7: 36-45. Simonsen JL, Rosada C, Serakinci N, et al. 2002. Telomerase expression extends the proliferative life-span and maintains the osteogenic potential of human bone marrow stromal cells. Nat Biotechnol 20: 592-596. Haynesworth SE, Baber MA, Caplan AI. 1995. Characterization of the unique mesenchymal stem cell phenotype in vitro. Trans 41st Ann Meet Orthop Res Soc 20: 7. Kratchmarova I, Blagoev B, Haack-Sorensen M, et al. 2005. Mechanism of divergent growth factor effects in mesenchymal stem cell differentiation. Science 308: 1472-1477. Liu L, DiGirolamo CM, Navarro PA, et al. 2004. Telomerase deficiency impairs differentiation of mesenchymal stem cells. Exp Cell Res 294: 1-8. Phinney DG, Kopen G, Righter W, et al. 1999. Donor variation in the growth properties and osteogenic potential of human marrow stromal cells. J Cell Biochem 75: 424-436. McBeath R, Pirone DM, Nelson CM, et al. 2004. Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev Cell 6: 483-495. Jochum W, David JP, Elliott C, et al. 2000. Increased bone formation and osteosclerosis in mice overexpressing the transcription factor Fra-1. Nat Med 6: 980-984. de Boer J, Siddappa R, Gaspar C, et al. 2004. Wnt signaling inhibits osteogenic differentiation of human mesenchymal stem cells. Bone 34: 818-826. Both SK, van der Muijsenberg AM, van Blitterswijk CA, et al. 2007. A rapid and labour efficient method for expansion of human mesenchymal stem cells for clinical applications. Tissue Eng 13: 3-9. Balooch G, Balooch M, Nalla RK, et al. 2005. TGF-beta regulates the mechanical properties and composition of bone matrix. Proc Natl Acad Sci USA 102: 18813-18818. Urist MR. 1965. Bone: formation by autoinduction. Science 150: 893-899. Jaiswal N, Haynesworth SE, Caplan AI, et al. 1997. Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro. J Cell Biochem 64: 295-312. Seiler JG 3rd, Johnson J. 2000. Iliac crest autogenous bone grafting: donor site complications. J South Orthop Assoc 9: 91-97. Sugiyama O, An DS, Kung SP, et al. 2005. Lentivirus-mediated gene transfer induces long-term transgene expression of BMP-2 in vitro and new bone formation in vivo. Mol Ther 11: 390-398. Kassem M, Ankersen L, Eriksen EF, et al. 1997. Demonstration of cellular aging and senescence in serially passaged long-term cultures of human trabecular osteoblasts. Osteoporos Int 7: 514-524. Hill NM, Horne JG, Devane PA. 1999. Donor site morbidity in the iliac crest bone graft. Aust N Z J Surg 69: 726-728. Bruder SP, Fox BS. 1999. Tissue engineering of bone. Cell based strategies. Clin Orthop Related Res S68-S83. Barry FP, Murphy JM. 2004. Mesenchymal stem cells: clinical applications and biological characterization. Int J Biochem Cell Biol 36: 568-584. Mundy G, Garrett R, Harris S, et al. 1999. Stimulation of bone formation in vitro and in rodents by statins. Science 286: 1946-1949. Boer JD, Licht R, Bongers M, et al. 2006. Inhibition of histone acetylation as a tool in bone tissue engineering. Tissue Eng 12: 2927-2937. Haynesworth SE, Baber MA, Caplan AI. 1992. Cell surface antigens on human marrow-derived mesenchymal cells are detected by monoclonal antibodies. Bone 13: 69-80. Gronthos S, Zannettino AC, Hay SJ, et al. 2003. Molecular and cellular characterisation of highly purified stromal stem cells derived from human bone marrow. J Cell Sci 116: 1827-1835. Jorgensen NR, Henriksen Z, Sorensen OH, et al. 2004. Dexamethasone, BMP-2, and 1,25-dihydroxyvitamin D enhance a more differentiated osteoblast phenotype: validation of an in vitro model for human bone marrow-derived primary osteoblasts. Steroids 69: 219-226. Pittenger MF, et al. 1999. Multilineage potential of adult human mesenchymal stem cells. Science 284: 143-147. Derubeis AR, Cancedda R. 2004. Bone marrow stromal cells (BMSCs) in bone engineering: limitations and recent advances. Ann Biomed Eng 32: 160-165. De Boer J, Wang HJ, Van Blitterswijk C. 2004. Effects of Wnt signaling on proliferation and differentiation of human mesenchymal stem cells. Tissue Eng 10: 393-401. Yuan H, Van Den Doel M, Li S, et al. 2002. A comparison of the osteoinductive potential of two calcium phosphate ceramics implanted intramuscularly in goats. J Mater Sci Mater Med 13: 1271-1275. Bauer TW, Muschler GF. 2000. Bone graft materials. An overview of the basic science. Clin Orthop Related Res 10-27. Pettway GJ, Schneider A, Koh AJ, et al. 2005. Anabolic actions of PTH (1-34): use of a novel tissue engineering model to investigate temporal effects on bone. Bone 36: 959-970. Oreffo RO, Bord S, Triffitt JT. 1998. Skeletal progenitor cells and ageing human populations. Clin Sci (Lond) 94: 549-555. Frank O, Heim M, Jakob M, et al. 2002. Real-time quantitative RT-PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro. J Cell Biochem 85: 737-746. Liu P, Oyajobi BO, Russell RG, et al. 1999. Regulation of osteogenic differentiation of human bone marrow stromal cells: interaction between transforming growth factor-beta and 1,25(OH)(2) vitamin D(3) In vitro. Calcif Tissue Int 65: 173-180. Jaiswal RK, Jaiswal N, Bruder SP, et al. 2000. Adult human mesenchymal stem cell differentiation to the osteogenic or adipogenic lineage is regulated by mitogen-activated protein kinase. J Biol Chem 275: 9645-9652. Koch H, Jadlowiec JA, Campbell PG. 2005. Insulin-like growth factor-I induces early osteoblast gene expression in human mesenchymal stem cells. Stem Cells Dev 14: 621-631. Martin I, Muraglia A, Campanile G, et al. 1997. Fibroblast growth factor-2 supports ex vivo expansion and maintenance of osteogenic precursors from human bone marrow. Endocrinology 138: 4456-4462. Kruyt MC, de Bruijn JD, Yuan H, et al. 2004. Optimization of bone tissue engineering in goats: a peroperative seeding method using cryopreserved cells and localized bone formation in calcium phosphate scaffolds. Transplantation 77: 359-365. Abdelaal MM, Tholpady SS, Kessler JD, et al. 2004. BMP-9-transduced prefabricated muscular flaps for the treatment of bony defects. J Craniofac Surg 15: 736-741; discussion 742-744. Bertone AL, Pittman DD, Bouxsein ML, et al. 2004. Adenoviral-mediated transfer of human BMP-6 gene accelerates healing in a rabbit ulnar osteotomy model. J Orthop Res 22: 1261-1270. Zernik J, Twarog K, Upholt WB. 1990. Regulation of alkaline phosphatase and alpha 2(I) procollagen synthesis during early intramembranous bone formation in the rat mandible. Differentiation 44: 207-215. Tezuka K, Yasuda M, Watanabe N, et al. 2002. Stimulation of osteoblastic cell differentiation by Notch. J Bone Miner Res 17: 231-239. Dezawa M, et al. 2005. Bone marrow stromal cells generate muscle cells and repair muscle degeneration. Science 309: 314-317. Hong JH, Hwang ES, McManus MT, et al. 2005. TAZ, a transcriptional modulator of mesenchymal stem cell differentiation. Science 309: 1074-1078. Xiaoxue Y, Zhongqiang C, Zhaoqing G, et al. 2004. Immortalization of human osteoblasts by transferring human telomerase reverse transcriptase gene. Biochem Biophys Res Commun 315: 643-651. Tsuda H, Wada T, Ito Y, et al. 2003. Efficient BMP2 gene transfer and bone formation of mesenchymal stem cells by a fiber-mutant adenoviral vector. Mol Ther 7: 354-365. Hench LL, Polak JM. 1965. Third-generation biomedical materials. Science 295: 1014-1017. Kveiborg M, Kassem M, Langdahl B, et al. 1999. Telomere shortening during aging of human osteoblasts in vitro and leukocytes in vivo: lack of excessive telomere loss in osteoporotic patients. Mech Ageing Dev 106: 261-271. Shi S, Gronthos S, Chen S, et al. 2002. Bone formation by human postnatal bone marrow stromal stem cells is enhanced by telomerase expression. Nat Biotechnol 20: 587-591. Bruder SP, Jaiswal N, Ricalton NS, et al. 1998. Mesenchymal stem cells in osteobiology and applied bone regeneration. Clin Orthop Relat Res S247-S256. Petite H, Viateau V, Bensaid W, et al. 2000. Tissue-engineered bone regeneration. Nat Biotechnol 18: 959-963. Mendes SC, Tibbe JM, Veenhof M, et al. 2004. Relation between in vitro and in vivo osteogenic potential of cultured human bone marrow stromal cells. J Mater Sci Mater Med 15: 1123-1128. Bruder SP, Horowitz MC, Mosca JD, et al. 1997. Monoclonal antibodies reactive with human osteogenic cell surface antigens. Bone 21: 225-235. Yu X, Botchwey EA, Levine EM, et al. 2004. Bioreactor-based bone tissue engineering: the influence of dynamic flow on osteoblast phenotypic expression and matrix mineralization. Proc Natl Acad Sci USA 101: 11203-11208. Harley CB, Futcher AB, Greider CW. 1990. Telomeres shorten during ageing of human fibroblasts. Nature 345: 458-460. 2004; 22 2002; 17 1990; 345 2003; 116 2000; 6 2004; 69 2000; 9 2002; 13 2002; 99 1999; 286 2004; 6 1999; 284 1992; 13 1997; 7 1995; 20 2004; 32 2004; 77 2000; 18 1990; 44 2002; 85 2000 2005; 102 2004; 36 1965; 150 2004; 294 2003; 7 2004; 34 1999; 14 2005; 308 2005; 309 1998; 94 2005; 36 2004; 101 1997; 138 2006; 12 1997; 21 1997; 64 1999; 69 1998 1999; 65 2000; 275 1999; 106 2001; 25 2007; 13 1999 2004; 10 2004; 11 2002; 20 2004; 15 1965; 295 2005; 7 1999; 75 2004; 315 2005; 11 2005; 12 2005; 14 e_1_2_1_41_2 e_1_2_1_22_2 e_1_2_1_45_2 e_1_2_1_60_2 e_1_2_1_20_2 e_1_2_1_43_2 e_1_2_1_62_2 e_1_2_1_26_2 e_1_2_1_49_2 e_1_2_1_24_2 e_1_2_1_47_2 e_1_2_1_28_2 e_1_2_1_6_2 e_1_2_1_54_2 e_1_2_1_56_2 e_1_2_1_2_2 e_1_2_1_12_2 e_1_2_1_33_2 e_1_2_1_50_2 e_1_2_1_10_2 e_1_2_1_31_2 e_1_2_1_16_2 e_1_2_1_37_2 e_1_2_1_14_2 e_1_2_1_35_2 e_1_2_1_58_2 e_1_2_1_8_2 e_1_2_1_18_2 e_1_2_1_39_2 e_1_2_1_40_2 e_1_2_1_23_2 e_1_2_1_44_2 e_1_2_1_61_2 e_1_2_1_21_2 e_1_2_1_42_2 e_1_2_1_27_2 e_1_2_1_48_2 e_1_2_1_25_2 e_1_2_1_46_2 e_1_2_1_29_2 Seiler JG (e_1_2_1_4_2) 2000; 9 e_1_2_1_30_2 e_1_2_1_53_2 e_1_2_1_7_2 e_1_2_1_55_2 e_1_2_1_5_2 e_1_2_1_11_2 e_1_2_1_34_2 e_1_2_1_3_2 e_1_2_1_32_2 e_1_2_1_51_2 e_1_2_1_15_2 e_1_2_1_38_2 e_1_2_1_13_2 e_1_2_1_36_2 Haynesworth SE (e_1_2_1_52_2) 1995; 20 e_1_2_1_19_2 e_1_2_1_57_2 e_1_2_1_17_2 e_1_2_1_59_2 e_1_2_1_9_2 |
| References_xml | – reference: Cowan CM, et al. 2004. Adipose-derived adult stromal cells heal critical-size mouse calvarial defects. Nat Biotechnol 22: 560-567. – reference: Kratchmarova I, Blagoev B, Haack-Sorensen M, et al. 2005. Mechanism of divergent growth factor effects in mesenchymal stem cell differentiation. Science 308: 1472-1477. – reference: Sugiyama O, An DS, Kung SP, et al. 2005. Lentivirus-mediated gene transfer induces long-term transgene expression of BMP-2 in vitro and new bone formation in vivo. Mol Ther 11: 390-398. – reference: Pittenger MF, et al. 1999. Multilineage potential of adult human mesenchymal stem cells. Science 284: 143-147. – reference: Livak KJ, Schmittgen TD. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25: 402-408. – reference: Yu X, Botchwey EA, Levine EM, et al. 2004. Bioreactor-based bone tissue engineering: the influence of dynamic flow on osteoblast phenotypic expression and matrix mineralization. Proc Natl Acad Sci USA 101: 11203-11208. – reference: Frank O, Heim M, Jakob M, et al. 2002. Real-time quantitative RT-PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro. J Cell Biochem 85: 737-746. – reference: Shi S, Gronthos S, Chen S, et al. 2002. Bone formation by human postnatal bone marrow stromal stem cells is enhanced by telomerase expression. Nat Biotechnol 20: 587-591. – reference: Le Blanc K, Pittenger M. 2005. Mesenchymal stem cells: progress toward promise. Cytotherapy 7: 36-45. – reference: Xiaoxue Y, Zhongqiang C, Zhaoqing G, et al. 2004. Immortalization of human osteoblasts by transferring human telomerase reverse transcriptase gene. Biochem Biophys Res Commun 315: 643-651. – reference: Liu P, Oyajobi BO, Russell RG, et al. 1999. Regulation of osteogenic differentiation of human bone marrow stromal cells: interaction between transforming growth factor-beta and 1,25(OH)(2) vitamin D(3) In vitro. Calcif Tissue Int 65: 173-180. – reference: Petite H, Viateau V, Bensaid W, et al. 2000. Tissue-engineered bone regeneration. Nat Biotechnol 18: 959-963. – reference: Pola E, Gao W, Zhou Y, et al. 2004. Efficient bone formation by gene transfer of human LIM mineralization protein-3. Gene Ther 11: 683-693. – reference: Bauer TW, Muschler GF. 2000. Bone graft materials. An overview of the basic science. Clin Orthop Related Res 10-27. – reference: Mendes SC, Tibbe JM, Veenhof M, et al. 2004. Relation between in vitro and in vivo osteogenic potential of cultured human bone marrow stromal cells. J Mater Sci Mater Med 15: 1123-1128. – reference: Bruder SP, Jaiswal N, Ricalton NS, et al. 1998. Mesenchymal stem cells in osteobiology and applied bone regeneration. Clin Orthop Relat Res S247-S256. – reference: Seiler JG 3rd, Johnson J. 2000. Iliac crest autogenous bone grafting: donor site complications. J South Orthop Assoc 9: 91-97. – reference: Haynesworth SE, Baber MA, Caplan AI. 1995. Characterization of the unique mesenchymal stem cell phenotype in vitro. Trans 41st Ann Meet Orthop Res Soc 20: 7. – reference: Bancroft GN, Sikavitsas VI, van den Dolder J, et al. 2002. Fluid flow increases mineralized matrix deposition in 3D perfusion culture of marrow stromal osteoblasts in a dose-dependent manner. Proc Natl Acad Sci USA 99: 12600-12605. – reference: Harley CB, Futcher AB, Greider CW. 1990. Telomeres shorten during ageing of human fibroblasts. Nature 345: 458-460. – reference: Urist MR. 1965. Bone: formation by autoinduction. Science 150: 893-899. – reference: Tezuka K, Yasuda M, Watanabe N, et al. 2002. Stimulation of osteoblastic cell differentiation by Notch. J Bone Miner Res 17: 231-239. – reference: Yuan H, Van Den Doel M, Li S, et al. 2002. A comparison of the osteoinductive potential of two calcium phosphate ceramics implanted intramuscularly in goats. J Mater Sci Mater Med 13: 1271-1275. – reference: Martin I, Muraglia A, Campanile G, et al. 1997. Fibroblast growth factor-2 supports ex vivo expansion and maintenance of osteogenic precursors from human bone marrow. Endocrinology 138: 4456-4462. – reference: Hill NM, Horne JG, Devane PA. 1999. Donor site morbidity in the iliac crest bone graft. Aust N Z J Surg 69: 726-728. – reference: Tsuda H, Wada T, Ito Y, et al. 2003. Efficient BMP2 gene transfer and bone formation of mesenchymal stem cells by a fiber-mutant adenoviral vector. Mol Ther 7: 354-365. – reference: Hong JH, Hwang ES, McManus MT, et al. 2005. TAZ, a transcriptional modulator of mesenchymal stem cell differentiation. Science 309: 1074-1078. – reference: Jochum W, David JP, Elliott C, et al. 2000. Increased bone formation and osteosclerosis in mice overexpressing the transcription factor Fra-1. Nat Med 6: 980-984. – reference: Derubeis AR, Cancedda R. 2004. Bone marrow stromal cells (BMSCs) in bone engineering: limitations and recent advances. Ann Biomed Eng 32: 160-165. – reference: Koch H, Jadlowiec JA, Campbell PG. 2005. Insulin-like growth factor-I induces early osteoblast gene expression in human mesenchymal stem cells. Stem Cells Dev 14: 621-631. – reference: Kassem M, Ankersen L, Eriksen EF, et al. 1997. Demonstration of cellular aging and senescence in serially passaged long-term cultures of human trabecular osteoblasts. Osteoporos Int 7: 514-524. – reference: Barry FP, Murphy JM. 2004. Mesenchymal stem cells: clinical applications and biological characterization. Int J Biochem Cell Biol 36: 568-584. – reference: Kruyt MC, de Bruijn JD, Yuan H, et al. 2004. Optimization of bone tissue engineering in goats: a peroperative seeding method using cryopreserved cells and localized bone formation in calcium phosphate scaffolds. Transplantation 77: 359-365. – reference: D'Ippolito G, Schiller PC, Ricordi C, et al. 1999. Age-related osteogenic potential of mesenchymal stromal stem cells from human vertebral bone marrow. J Bone Miner Res 14: 1115-1122. – reference: Jaiswal RK, Jaiswal N, Bruder SP, et al. 2000. Adult human mesenchymal stem cell differentiation to the osteogenic or adipogenic lineage is regulated by mitogen-activated protein kinase. J Biol Chem 275: 9645-9652. – reference: Pettway GJ, Schneider A, Koh AJ, et al. 2005. Anabolic actions of PTH (1-34): use of a novel tissue engineering model to investigate temporal effects on bone. Bone 36: 959-970. – reference: Zernik J, Twarog K, Upholt WB. 1990. Regulation of alkaline phosphatase and alpha 2(I) procollagen synthesis during early intramembranous bone formation in the rat mandible. Differentiation 44: 207-215. – reference: Kveiborg M, Kassem M, Langdahl B, et al. 1999. Telomere shortening during aging of human osteoblasts in vitro and leukocytes in vivo: lack of excessive telomere loss in osteoporotic patients. Mech Ageing Dev 106: 261-271. – reference: Abdelaal MM, Tholpady SS, Kessler JD, et al. 2004. BMP-9-transduced prefabricated muscular flaps for the treatment of bony defects. J Craniofac Surg 15: 736-741; discussion 742-744. – reference: Fehrer C, Lepperdinger G. 2005. Mesenchymal stem cell aging. Exp Gerontol 12: 926-930. – reference: Liu L, DiGirolamo CM, Navarro PA, et al. 2004. Telomerase deficiency impairs differentiation of mesenchymal stem cells. Exp Cell Res 294: 1-8. – reference: Bruder SP, Jaiswal N, Haynesworth SE. 1997. Growth kinetics, self-renewal, and the osteogenic potential of purified human mesenchymal stem cells during extensive subcultivation and following cryopreservation. 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| SubjectTerms | Acetabulum - cytology Adult Aged Aged, 80 and over Alkaline Phosphatase - analysis Alkaline Phosphatase - genetics Animals Bone Substitutes bone tissue engineering Cell Differentiation - drug effects Dexamethasone - pharmacology Female Gene Expression Profiling Genetic Variation human mesenchymal stem cells Humans Ilium - cytology Male Mesenchymal Stromal Cells - cytology Mesenchymal Stromal Cells - physiology Mice Middle Aged Multipotent Stem Cells - cytology Multipotent Stem Cells - physiology Osteogenesis Tissue Donors - classification Tissue Engineering - methods |
| Title | Donor variation and loss of multipotency during in vitro expansion of human mesenchymal stem cells for bone tissue engineering |
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