Please use this identifier to cite or link to this item:
Effects of quantum dots and functional RGD-chimeric protein on the properties and differentiation of adipose-derived adult stem cells in an alginate three-dimensional culture system
Adult adipose-derived stem cells
Gap junction intercellular communication
Alginate three-dimensional culture
|引用:||1. Walling MA, Novak JA, Shepard JR. Quantum dots for live cell and in vivo imaging. Int J Mol Sci. 2009;10:441-491. 2. Tokumasu F, Fairhurst RM, Ostera GR, et al. Band 3 modifications in Plasmodium falciparum-infected AA and CC erythrocytes assayed by autocorrelation analysis using quantum dots. J Cell Sci. 2005;118:1091-1098. 3. Dahan M, Levi S, Luccardini C, et al. Diffusion dynamics of glycine receptors revealed by single-quantum dot tracking. Science. 2003;302:442-445. 4. Ballou B, Lagerholm BC, Ernst LA, et al. Noninvasive imaging of quantum dots in mice. Bioconjug Chem. 2004;15:79-86. 5. Jaiswal JK, Mattoussi H, Mauro JM, et al. Long-term multiple color imaging of live cells using quantum dot bioconjugates. Nat Biotechnol. 2003;21:47-51. 6. Henriques ST, Castanho MA. Environmental factors that enhance the action of the cell penetrating peptide pep-1 A spectroscopic study using lipidic vesicles. Biochim Biophys Acta. 2005;1669:75-86. 7. Hsieh SC, Wang FF, Hung SC, et al. The internalized CdSe/ZnS quantum dots impair the chondrogenesis of bone marrow mesenchymal stem cells. J Biomed Mater Res B Appl Biomater. 2006. 8. Hoshino A, Manabe N, Fujioka K, et al. Use of fluorescent quantum dot bioconjugates for cellular imaging of immune cells, cell organelle labeling, and nanomedicine: surface modification regulates biological function, including cytotoxicity. J Artif Organs. 2007;10:149-157. 9. Wong RC, Pera MF, Pebay A. Role of gap junctions in embryonic and somatic stem cells. Stem Cell Rev. 2008;4:283-292. 10. Lin TM, Chang HW, Wang KH, et al. Isolation and identification of mesenchymal stem cells from human lipoma tissue. Biochem Biophys Res Commun. 2007;361:883-889. 11. Matic M, Evans WH, Brink PR, et al. Epidermal stem cells do not communicate through gap junctions. J Invest Dermatol. 2002;118:110-116. 12. Oviedo NJ, Levin M. smedinx-11 is a planarian stem cell gap junction gene required for regeneration and homeostasis. Development. 2007;134:3121-3131. 13. Stains JP, Civitelli R. Gap junctions in skeletal development and function. Biochim Biophys Acta. 2005;1719:69-81. 14. Jen AC, Wake MC, Mikos AG. Review: Hydrogels for cell immobilization. Biotechnol Bioeng. 1996;50:357-364. 15. Martinsen A, Skjak-Braek G, Smidsrod O. Alginate as immobilization material: I. Correlation between chemical and physical properties of alginate gel beads. Biotechnol Bioeng. 1989;33:79-89. 16. Smetana K, Jr. Cell biology of hydrogels. Biomaterials. 1993;14:1046-1050. 17. Ostuni E, Whitesides GM, Ingber DE, et al. Using self-assembled monolayers to pattern ECM proteins and cells on substrates. Methods Mol Biol. 2009;522:183-194. 18. Connelly JT, Garcia AJ, Levenston ME. Inhibition of in vitro chondrogenesis in RGD-modified three-dimensional alginate gels. Biomaterials. 2007;28:1071-1083. 19. Salinas CN, Anseth KS. The enhancement of chondrogenic differentiation of human mesenchymal stem cells by enzymatically regulated RGD functionalities. Biomaterials. 2008;29:2370-2377. 20. Wee S, Gombotz WR. Protein release from alginate matrices. Adv Drug Deliv Rev. 1998;31:267-285. 21. Smidsrod O, Skjak-Braek G. Alginate as immobilization matrix for cells. Trends Biotechnol. 1990;8:71-78. 22. Giuliano KA, Post PL, Hahn KM, et al. Fluorescent protein biosensors: measurement of molecular dynamics in living cells. Annu Rev Biophys Biomol Struct. 1995;24:405-434. 23. Gao X, Cui Y, Levenson RM, et al. In vivo cancer targeting and imaging with semiconductor quantum dots. Nat Biotechnol. 2004;22:969-976. 24. Sukhanova A, Devy J, Venteo L, et al. Biocompatible fluorescent nanocrystals for immunolabeling of membrane proteins and cells. Anal Biochem. 2004;324:60-67. 25. Finley KR, Davidson AE, Ekker SC. Three-color imaging using fluorescent proteins in living zebrafish embryos. Biotechniques. 2001;31:66-70, 72. 26. Medintz IL, Uyeda HT, Goldman ER, et al. Quantum dot bioconjugates for imaging, labelling and sensing. Nat Mater. 2005;4:435-446. 27. Chan WC, Maxwell DJ, Gao X, et al. Luminescent quantum dots for multiplexed biological detection and imaging. Curr Opin Biotechnol. 2002;13:40-46. 28. Zhang Y, So MK, Rao J. Protease-modulated cellular uptake of quantum dots. Nano Lett. 2006;6:1988-1992. 29. Cai W, Shin DW, Chen K, et al. Peptide-labeled near-infrared quantum dots for imaging tumor vasculature in living subjects. Nano Lett. 2006;6:669-676. 30. Delehanty JB, Medintz IL, Pons T, et al. Self-assembled quantum dot-peptide bioconjugates for selective intracellular delivery. Bioconjug Chem. 2006;17:920-927. 31. Voura EB, Jaiswal JK, Mattoussi H, et al. Tracking metastatic tumor cell extravasation with quantum dot nanocrystals and fluorescence emission-scanning microscopy. Nat Med. 2004;10:993-998. 32. Chan WC, Nie S. Quantum dot bioconjugates for ultrasensitive nonisotopic detection. Science. 1998;281:2016-2018. 33. Dubertret B, Skourides P, Norris DJ, et al. In vivo imaging of quantum dots encapsulated in phospholipid micelles. Science. 2002;298:1759-1762. 34. Hsieh SC, Wang FF, Lin CS, et al. The inhibition of osteogenesis with human bone marrow mesenchymal stem cells by CdSe/ZnS quantum dot labels. Biomaterials. 2006;27:1656-1664. 35. Baddoo M, Hill K, Wilkinson R, et al. Characterization of mesenchymal stem cells isolated from murine bone marrow by negative selection. J Cell Biochem. 2003;89:1235-1249. 36. Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potential of adult human mesenchymal stem cells. Science. 1999;284:143-147. 37. Ceresa BP, Schmid SL. Regulation of signal transduction by endocytosis. Curr Opin Cell Biol. 2000;12:204-210. 38. Hayes S, Chawla A, Corvera S. TGF beta receptor internalization into EEA1-enriched early endosomes: role in signaling to Smad2. J Cell Biol. 2002;158:1239-1249. 39. Lundberg P, Langel U. A brief introduction to cell-penetrating peptides. J Mol Recognit. 2003;16:227-233. 40. Gros E, Deshayes S, Morris MC, et al. A non-covalent peptide-based strategy for protein and peptide nucleic acid transduction. Biochim Biophys Acta. 2006. 41. Morris MC, Depollier J, Mery J, et al. A peptide carrier for the delivery of biologically active proteins into mammalian cells. Nat Biotechnol. 2001;19:1173-1176. 42. Mattheakis LC, Dias JM, Choi YJ, et al. Optical coding of mammalian cells using semiconductor quantum dots. Anal Biochem. 2004;327:200-208. 43. Seleverstov O, Zabirnyk O, Zscharnack M, et al. Quantum dots for human mesenchymal stem cells labeling. A size-dependent autophagy activation. Nano Lett. 2006;6:2826-2832. 44. Slotkin JR, Chakrabarti L, Dai HN, et al. In vivo quantum dot labeling of mammalian stem and progenitor cells. Dev Dyn. 2007;236:3393-3401. 45. Manabe N, Hoshino A, Liang YQ, et al. Quantum dot as a drug tracer in vivo. IEEE Trans Nanobioscience. 2006;5:263-267. 46. Shah BS, Clark PA, Moioli EK, et al. Labeling of mesenchymal stem cells by bioconjugated quantum dots. Nano Lett. 2007;7:3071-3079. 47. Chang JC, Su HL, Hsu SH. The use of peptide-delivery to protect human adipose-derived adult stem cells from damage caused by the internalization of quantum dots. Biomaterials. 2008;29:925-936. 48. Lidke DS, Nagy P, Jovin TM, et al. Biotin-ligand complexes with streptavidin quantum dots for in vivo cell labeling of membrane receptors. Methods Mol Biol. 2007;374:69-79. 49. Seleverstov O, Zabirnyk O, Zscharnack M, et al. Quantum dots for human mesenchymal stem cells labeling. A size-dependent autophagy activation. Nano Lett. 2006;6:2826-2832. 50. Gupta AK, Curtis AS. Lactoferrin and ceruloplasmin derivatized superparamagnetic iron oxide nanoparticles for targeting cell surface receptors. Biomaterials. 2004;25:3029-3040. 51. Gupta AK, Gupta M, Yarwood SJ, et al. Effect of cellular uptake of gelatin nanoparticles on adhesion, morphology and cytoskeleton organisation of human fibroblasts. J Control Release. 2004;95:197-207. 52. Liu CZ, Wang YW, Shen MC, et al. Analysis of human platelet glycoprotein IIb-IIIa by fluorescein isothiocyanate-conjugated disintegrins with flow cytometry. Thromb Haemost. 1994;72:919-925. 53. Hsieh SC, Wang FF, Hung SC, et al. The internalized CdSe/ZnS quantum dots impair the chondrogenesis of bone marrow mesenchymal stem cells. J Biomed Mater Res B Appl Biomater. 2006. 54. Schroeder JE, Shweky I, Shmeeda H, et al. Folate-mediated tumor cell uptake of quantum dots entrapped in lipid nanoparticles. J Control Release. 2007;124:28-34. 55. Hardman R. A toxicologic review of quantum dots: toxicity depends on physicochemical and environmental factors. Environ Health Perspect. 2006;114:165-172. 56. Stains JP, Civitelli R. Gap junctions in skeletal development and function. Biochim Biophys Acta. 2005;1719:69-81. 57. Houghton FD. Role of gap junctions during early embryo development. Reproduction. 2005;129:129-135. 58. Laird DW, Puranam KL, Revel JP. Turnover and phosphorylation dynamics of connexin43 gap junction protein in cultured cardiac myocytes. Biochem J. 1991;273(Pt 1):67-72. 59. Lampe PD, Lau AF. Regulation of gap junctions by phosphorylation of connexins. Arch Biochem Biophys. 2000;384:205-215. 60. Evans WH, Martin PE. Gap junctions: structure and function (Review). Mol Membr Biol. 2002;19:121-136. 61. Valiunas V, Doronin S, Valiuniene L, et al. Human mesenchymal stem cells make cardiac connexins and form functional gap junctions. J Physiol. 2004;555:617-626. 62. Worsdorfer P, Maxeiner S, Markopoulos C, et al. Connexin expression and functional analysis of gap junctional communication in mouse embryonic stem cells. Stem Cells. 2008;26:431-439. 63. Trosko JE, Chang CC, Wilson MR, et al. Gap junctions and the regulation of cellular functions of stem cells during development and differentiation. Methods. 2000;20:245-264. 64. Mauck RL, Byers BA, Yuan X, et al. Regulation of cartilaginous ECM gene transcription by chondrocytes and MSCs in 3D culture in response to dynamic loading. Biomech Model Mechanobiol. 2007;6:113-125. 65. Meinel L, Hofmann S, Karageorgiou V, et al. Engineering cartilage-like tissue using human mesenchymal stem cells and silk protein scaffolds. Biotechnol Bioeng. 2004;88:379-391. 66. Hwang NS, Varghese S, Zhang Z, et al. Chondrogenic differentiation of human embryonic stem cell-derived cells in arginine-glycine-aspartate-modified hydrogels. Tissue Eng. 2006;12:2695-2706. 67. Yamaoka H, Asato H, Ogasawara T, et al. Cartilage tissue engineering using human auricular chondrocytes embedded in different hydrogel materials. J Biomed Mater Res A. 2006;78:1-11. 68. Chen DC, Hu NT, Chen Y.J., et al. Chimeric Proteins with a Cellulose Binding Domain. 2002:6-18. 69. Shoseyov O, Warren RAJ. Cellulose binding domains-A novel fusion technology for efficient, low cost purification and immobilization of recombinant proteins. inNovations. 1997;7:1-3. 70. Hsu SH, Chu WP, Lin YS, et al. The effect of an RGD-containing fusion protein CBD-RGD in promoting cellular adhesion. J Biotechnol. 2004;111:143-154. 71. Hsu SH, Chuang SC, Chen CH, et al. Endothelial cell attachment to the gamma irradiated small diameter polyurethane vascular grafts. Biomed Mater Eng. 2006;16:397-404. 72. Hsu SH, Sun SH, Chen DC. Improved retention of endothelial cells seeded on polyurethane small-diameter vascular grafts modified by a recombinant RGD-containing protein. Artif Organs. 2003;27:1068-1078. 73. Hsu SH, Whu SW, Hsieh SC, et al. Evaluation of chitosan-alginate-hyaluronate complexes modified by an RGD-containing protein as tissue-engineering scaffolds for cartilage regeneration. Artif Organs. 2004;28:693-703. 74. Tavella S, Bellese G, Castagnola P, et al. Regulated expression of fibronectin, laminin and related integrin receptors during the early chondrocyte differentiation. J Cell Sci. 1997;110 ( Pt 18):2261-2270. 75. DeLise AM, Fischer L, Tuan RS. Cellular interactions and signaling in cartilage development. Osteoarthritis Cartilage. 2000;8:309-334. 76. Cali G, Mazzarella C, Chiacchio M, et al. RhoA activity is required for fibronectin assembly and counteracts beta 1B integrin inhibitory effect in FRT epithelial cells. Journal of Cell Science. 1999;112:957-965. 77. Defilippi P, Venturino M, Gulino D, et al. Dissection of pathways implicated in integrin-mediated actin cytoskeleton assembly - Involvement of protein kinase C, Rho GTPase, and tyrosine phosphorylation. Journal of Biological Chemistry. 1997;272:21726-21734. 78. Parsons JT. Integrin-mediated signalling: Regulation by protein tyrosine kinases and small GTP-binding proteins. Current Opinion in Cell Biology. 1996;8:146-152. 79. Ridley AJ, Hall A. The Small Gtp-Binding Protein Rho Regulates the Assembly of Focal Adhesions and Actin Stress Fibers in Response to Growth-Factors. Cell. 1992;70:389-399. 80. Lee J, Ko M, Joo CK. Rho plays a key role in TGF-beta1-induced cytoskeletal rearrangement in human retinal pigment epithelium. J Cell Physiol. 2008;216:520-526. 81. Woods A, Wang G, Beier F. RhoA/ROCK signaling regulates Sox9 expression and actin organization during chondrogenesis. J Biol Chem. 2005;280:11626-11634. 82. Woods A, Beier F. RhoA/ROCK signaling regulates chondrogenesis in a context-dependent manner. J Biol Chem. 2006;281:13134-13140. 83. Wickham MQ, Erickson GR, Gimble JM, et al. Multipotent stromal cells derived from the infrapatellar fat pad of the knee. Clin Orthop Relat Res. 2003;196-212. 84. Zuk PA, Zhu M, Ashjian P, et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell. 2002;13:4279-4295. 85. Liu CZ, Wang YW, Shen MC, et al. Analysis of human platelet glycoprotein IIb-IIIa by fluorescein isothiocyanate-conjugated disintegrins with flow cytometry. Thromb Haemost. 1994;72:919-925. 86. Laird DW. Connexin phosphorylation as a regulatory event linked to gap junction internalization and degradation. Biochim Biophys Acta. 2005;1711:172-182. 87. Rosowski M, Falb M, Tschirschmann M, et al. Initiation of mesenchymal condensation in alginate hollow spheres--a useful model for understanding cartilage repair? Artif Organs. 2006;30:775-784. 88. Wade MH, Trosko JE, Schindler M. A fluorescence photobleaching assay of gap junction-mediated communication between human cells. Science. 1986;232:525-528. 89. Hanaki K, Momo A, Oku T, et al. Semiconductor quantum dot/albumin complex is a long-life and highly photostable endosome marker. Biochem Biophys Res Commun. 2003;302:496-501. 90. Hirschey MD, Han YJ, Stucky GD, et al. Imaging Escherichia coli using functionalized core/shell CdSe/CdS quantum dots. J Biol Inorg Chem. 2006;11:663-669. 91. Kriete A, Papazoglou E, Edrissi B, et al. Automated quantification of quantum-dot-labelled epidermal growth factor receptor internalization via multiscale image segmentation. J Microsc. 2006;222:22-27. 92. Lewin M, Carlesso N, Tung CH, et al. Tat peptide-derivatized magnetic nanoparticles allow in vivo tracking and recovery of progenitor cells. Nat Biotechnol. 2000;18:410-414. 93. Lovric J, Bazzi HS, Cuie Y, et al. Differences in subcellular distribution and toxicity of green and red emitting CdTe quantum dots. J Mol Med. 2005;83:377-385. 94. Shiohara A, Hoshino A, Hanaki K, et al. On the cyto-toxicity caused by quantum dots. Microbiol Immunol. 2004;48:669-675. 95. Deshayes S, Morris MC, Divita G, et al. Interactions of primary amphipathic cell penetrating peptides with model membranes: consequences on the mechanisms of intracellular delivery of therapeutics. Curr Pharm Des. 2005;11:3629-3638. 96. Deshayes S, Heitz A, Morris MC, et al. Insight into the mechanism of internalization of the cell-penetrating carrier peptide Pep-1 through conformational analysis. Biochemistry. 2004;43:1449-1457. 97. Morris MC, Vidal P, Chaloin L, et al. A new peptide vector for efficient delivery of oligonucleotides into mammalian cells. Nucleic Acids Res. 1997;25:2730-2736. 98. Howe CL, Valletta JS, Rusnak AS, et al. NGF signaling from clathrin-coated vesicles: evidence that signaling endosomes serve as a platform for the Ras-MAPK pathway. Neuron. 2001;32:801-814. 99. Tang MX, Redemann CT, Szoka FC, Jr. In vitro gene delivery by degraded polyamidoamine dendrimers. Bioconjug Chem. 1996;7:703-714. 100. Terrone D, Sang SL, Roudaia L, et al. Penetratin and related cell-penetrating cationic peptides can translocate across lipid bilayers in the presence of a transbilayer potential. Biochemistry. 2003;42:13787-13799. 101. Dewar H, Warren DT, Gardiner FC, et al. Novel proteins linking the actin cytoskeleton to the endocytic machinery in Saccharomyces cerevisiae. Mol Biol Cell. 2002;13:3646-3661. 102. Loty S, Foll C, Forest N, et al. Association of enhanced expression of gap junctions with in vitro chondrogenic differentiation of rat nasal septal cartilage-released cells following their dedifferentiation and redifferentiation. Arch Oral Biol. 2000;45:843-856. 103. Zhang W, Green C, Stott NS. Bone morphogenetic protein-2 modulation of chondrogenic differentiation in vitro involves gap junction-mediated intercellular communication. J Cell Physiol. 2002;193:233-243. 104. Pampaloni F, Reynaud EG, Stelzer EH. The third dimension bridges the gap between cell culture and live tissue. Nat Rev Mol Cell Biol. 2007;8:839-845. 105. Lagerholm BC. Peptide-mediated intracellular delivery of quantum dots. Methods Mol Biol. 2007;374:105-112. 106. Dewar H, Warren DT, Gardiner FC, et al. Novel proteins linking the actin cytoskeleton to the endocytic machinery in Saccharomyces cerevisiae. Mol Biol Cell. 2002;13:3646-3661. 107. Gupta AK, Gupta M. Cytotoxicity suppression and cellular uptake enhancement of surface modified magnetic nanoparticles. Biomaterials. 2005;26:1565-1573. 108. Lin TM, Tsai JL, Lin SD, et al. Accelerated growth and prolonged lifespan of adipose tissue-derived human mesenchymal stem cells in a medium using reduced calcium and antioxidants. Stem Cells Dev. 2005;14:92-102. 109. Qin H, Shao Q, Igdoura SA, et al. Lysosomal and proteasomal degradation play distinct roles in the life cycle of Cx43 in gap junctional intercellular communication-deficient and -competent breast tumor cells. J Biol Chem. 2003;278:30005-30014. 110. Girao H, Pereira P. Phosphorylation of connexin 43 acts as a stimuli for proteasome-dependent degradation of the protein in lens epithelial cells. Mol Vis. 2003;9:24-30. 111. Larsen WJ, Tung HN, Murray SA, et al. Evidence for the participation of actin microfilaments and bristle coats in the internalization of gap junction membrane. J Cell Biol. 1979;83:576-587. 112. Cooper CD, Lampe PD. Casein kinase 1 regulates connexin-43 gap junction assembly. J Biol Chem. 2002;277:44962-44968. 113. Wert SE, Larsen WJ. Meiotic resumption and gap junction modulation in the cultured rat cumulus-oocyte complex. Gamete Res. 1989;22:143-162. 114. Johnson RG, Meyer RA, Li XR, et al. Gap junctions assemble in the presence of cytoskeletal inhibitors, but enhanced assembly requires microtubules. Exp Cell Res. 2002;275:67-80. 115. Murray SA, Williams SY, Dillard CY, et al. Relationship of cytoskeletal filaments to annular gap junction expression in human adrenal cortical tumor cells in culture. Exp Cell Res. 1997;234:398-404. 116. Juliano RL, Reddig P, Alahari S, et al. Integrin regulation of cell signalling and motility. Biochem Soc Trans. 2004;32:443-446. 117. Juliano RL. Signal transduction by cell adhesion receptors and the cytoskeleton: functions of integrins, cadherins, selectins, and immunoglobulin-superfamily members. Annu Rev Pharmacol Toxicol. 2002;42:283-323. 118. Presley CA, Lee AW, Kastl B, et al. Bone marrow connexin-43 expression is critical for hematopoietic regeneration after chemotherapy. Cell Commun Adhes. 2005;12:307-317. 119. Waterman-Storer CM, Salmon WC, Salmon ED. Feedback interactions between cell-cell adherens junctions and cytoskeletal dynamics in newt lung epithelial cells. Mol Biol Cell. 2000;11:2471-2483. 120. Ren XD, Kiosses WB, Schwartz MA. Regulation of the small GTP-binding protein Rho by cell adhesion and the cytoskeleton. EMBO J. 1999;18:578-585. 121. Stains JP, Lecanda F, Screen J, et al. Gap junctional communication modulates gene transcription by altering the recruitment of Sp1 and Sp3 to connexin-response elements in osteoblast promoters. J Biol Chem. 2003;278:24377-24387. 122. Loty S, Foll C, Forest N, et al. Association of enhanced expression of gap junctions with in vitro chondrogenic differentiation of rat nasal septal cartilage-released cells following their dedifferentiation and redifferentiation. Arch Oral Biol. 2000;45:843-856. 123. Trosko JE, Chang CC. Mechanism of up-regulated gap junctional intercellular communication during chemoprevention and chemotherapy of cancer. Mutat Res. 2001;480-481:219-229. 124. Trosko JE, Tai MH. Adult stem cell theory of the multi-stage, multi-mechanism theory of carcinogenesis: role of inflammation on the promotion of initiated stem cells. Contrib Microbiol. 2006;13:45-65. 125. Tai MH, Olson LK, Madhukar BV, et al. Characterization of gap junctional intercellular communication in immortalized human pancreatic ductal epithelial cells with stem cell characteristics. Pancreas. 2003;26:e18-e26. 126. Dowling-Warriner CV, Trosko JE. Induction of gap junctional intercellular communication, connexin43 expression, and subsequent differentiation in human fetal neuronal cells by stimulation of the cyclic AMP pathway. Neuroscience. 2000;95:859-868. 127. Gaustad KG, Boquest AC, Anderson BE, et al. Differentiation of human adipose tissue stem cells using extracts of rat cardiomyocytes. Biochem Biophys Res Commun. 2004;314:420-427. 128. Loewenstein WR, Rose B. The cell-cell channel in the control of growth. Semin Cell Biol. 1992;3:59-79. 129. Wilson MR, Close TW, Trosko JE. Cell population dynamics (apoptosis, mitosis, and cell-cell communication) during disruption of homeostasis. Exp Cell Res. 2000;254:257-268. 130. Trosko JE, Goodman JI. Intercellular communication may facilitate apoptosis: implications for tumor promotion. Mol Carcinog. 1994;11:8-12. 131. Bursch W, Oberhammer F, Schulte-Hermann R. Cell death by apoptosis and its protective role against disease. Trends Pharmacol Sci. 1992;13:245-251. 132. Binetruy B, Heasley L, Bost F, et al. Concise review: regulation of embryonic stem cell lineage commitment by mitogen-activated protein kinases. Stem Cells. 2007;25:1090-1095. 133. Lee JW, Kim YH, Kim SH, et al. Chondrogenic differentiation of mesenchymal stem cells and its clinical applications. Yonsei Med J. 2004;45 Suppl:41-47. 134. Wilkins MR, Gasteiger E, Bairoch A, et al. Protein identification and analysis tools in the ExPASy server. Methods Mol Biol. 1999;112:531-552. 135. Mardilovich A, Craig JA, McCammon MQ, et al. Design of a novel fibronectin-mimetic peptide-amphiphile for functionalized biomaterials. Langmuir. 2006;22:3259-3264. 136. Garcia AJ, Schwarzbauer JE, Boettiger D. Distinct activation states of alpha5beta1 integrin show differential binding to RGD and synergy domains of fibronectin. Biochemistry. 2002;41:9063-9069. 137. Leahy DJ, Aukhil I, Erickson HP. 2.0 A crystal structure of a four-domain segment of human fibronectin encompassing the RGD loop and synergy region. Cell. 1996;84:155-164. 138. Dillo AK, Ochsenhirt SE, McCarthy JB, et al. Adhesion of alpha5beta1 receptors to biomimetic substrates constructed from peptide amphiphiles. Biomaterials. 2001;22:1493-1505. 139. Connelly JT, Garcia AJ, Levenston ME. Interactions between integrin ligand density and cytoskeletal integrity regulate BMSC chondrogenesis. J Cell Physiol. 2008;217:145-154. 140. Puolakkainen PA, Ranchalis JE, Gombotz WR, et al. Novel delivery system for inducing quiescence in intestinal stem cells in rats by transforming growth factor beta 1. Gastroenterology. 1994;107:1319-1326. 141. Segi N, Yotsuyangi T, Ikeda K. Interaction of calcium-induced alginate beads with propranolol. Chem Pharm Bull. 1989;37:3092-3095. 142. Sawhney AS, Hubbell JA. Poly(ethylene oxide)-graft-poly(L-lysine) copolymers to enhance the biocompatibility of poly(L-lysine)-alginate microcapsule membranes. Biomaterials. 1992;13:863-870. 143. Petrie TA, Capadona JR, Reyes CD, et al. Integrin specificity and enhanced cellular activities associated with surfaces presenting a recombinant fibronectin fragment compared to RGD supports. Biomaterials. 2006;27:5459-5470. 144. Keselowsky BG, Collard DM, Garcia AJ. Integrin binding specificity regulates biomaterial surface chemistry effects on cell differentiation. Proc Natl Acad Sci U S A. 2005;102:5953-5957. 145. Watanabe H, de Caestecker MP, Yamada Y. Transcriptional cross-talk between Smad, ERK1/2, and p38 mitogen-activated protein kinase pathways regulates transforming growth factor-beta-induced aggrecan gene expression in chondrogenic ATDC5 cells. J Biol Chem. 2001;276:14466-14473. 146. Drissi MH, Li X, Sheu TJ, et al. Runx2/Cbfa1 stimulation by retinoic acid is potentiated by BMP2 signaling through interaction with Smad1 on the collagen X promoter in chondrocytes. J Cell Biochem. 2003;90:1287-1298. 147. Han F, Gilbert JR, Harrison G, et al. Transforming growth factor-beta1 regulates fibronectin isoform expression and splicing factor SRp40 expression during ATDC5 chondrogenic maturation. Exp Cell Res. 2007;313:1518-1532. 148. Song JJ, Aswad R, Kanaan RA, et al. Connective tissue growth factor (CTGF) acts as a downstream mediator of TGF-beta1 to induce mesenchymal cell condensation. J Cell Physiol. 2007;210:398-410. 149. Lu ZF, Zandieh DB, Huang CL, et al. Beta1 integrins regulate chondrogenesis and rock signaling in adipose stem cells. Biochem Biophys Res Commun. 2008;372:547-552. 150. Klein J, Stock J, Vorlop KD. Pore-Size and Properties of Spherical Ca-Alginate Biocatalysts. European Journal of Applied Microbiology and Biotechnology. 1983;18:86-91. 151. Rasmussen MR, Snabe T, Pedersen LH. Numerical modelling of insulin and amyloglucosidase release from swelling Ca-alginate beads. J Control Release. 2003;91:395-405. 152. Gombotz WR, Wee SF. Protein release from alginate matrices. Advanced Drug Delivery Reviews. 1998;31:267-285. 153. Mumper RJ, Hoffman AS, Puolakkainen PA, et al. Calcium-Alginate Beads for the Oral Delivery of Transforming Growth Factor-Beta(1) (Tgf-Beta(1)) - Stabilization of Tgf-Beta(1) by the Addition of Polyacrylic-Acid Within Acid-Treated Beads. Journal of Controlled Release. 1994;30:241-251. 154. Smidsrod O. Relative Extension of Alginates Having Different Chemical Composition. Carbohydrate Research. 1973;27:107-118.|
|摘要:||活體追蹤幹細胞技術能有助於釐清細胞於體內修復之機轉， 提升動物實驗之精準度與成效，但利用量子點 (CdSe/ZnS Quantum dots) 標定細胞已證實會抑制人類之骨髓幹細胞分化，其機轉未明。本研究將比較不同途徑運，Pep-1胜肽與PolyFect轉殖，運送10 nM量子點 (655 nm) 標定人類脂肪幹細胞 (human adipose-derivated adult stem cells) ，探討其對細胞標定成效、細胞特性與分化能力之影響。研究發現，相較於量子點轉殖，細胞標定效率並無顯著差異(98.2-94.4%)，但Pep-1運送會促進細胞質內量子點聚集，顯著提升螢光強度，儘管Pep-1運送之量子點容易隨細胞分裂而釋出或分散，但經過二十八天培養，其螢光強度仍顯著高於轉殖組約三倍；此外，Pep-1可促使量子點攝入後一天逃離溶小體 (lysosome) 分解途徑(endocytosis-independent pathway)，但相反地，PolyFect轉殖則會顯著增加溶小體表現，但經過二十一天培養，不論經由何種途徑運送，量子點均會於溶小體代謝；Pep-1運送能改善量子點經由轉殖攝入對細胞特性抑制之影響，包含維持表面標誌蛋白(CD29 和CD105) 、細胞增生與維生素D (1,25 dihydroxyvitamine D) 和轉化生長子(Transforming growth factor β3, TGF-β3) 所誘導之硬軟骨分化。推測其機轉可能與早期胞內量子點降解途徑有關， 因於溶小體內之量子點可能阻礙的胞內核內體/溶小體途徑 (endo-/lysome pathway) 所調控之訊息傳遞。
本研究同時發現，Pep-1運送之量子點會經由同時活化細胞骨架蛋白 (F-actin) 和增加extracellular signal-regulated kinase (ERK) 激酶磷酸化，促進43型接合素 (Connexin 43, Cx43) 磷酸化而導致蛋白降解，進而抑制Cx43所調控之細胞間縫隙連接(Cx43-mediated gap junction intracellular communication, GJIC) ；然而，以高密度三維培養Pep-1運送量子點標定之細胞七天，原先受抑制之縫隙連接 (gap junction) 則會由於 Cx43蛋白質合成顯著增加而促使GJIC功能恢復；此外，轉殖之量子點會抑制細胞分化但並不影響細胞Cx43蛋白與gap junction表現。儘管gap junction在幹細胞的調控角色仍未釐清，然而多數研究證實原始未分化之間葉幹細胞僅表現少量Cx43和gap junction，本研究結果推測暫時性抑制GJIC可能有助於減少量子點攝入所導致的細胞毒性。
褐藻膠(alginate hydrogel)具有可降解與良好生物相容特性，已廣泛應用於藥物釋放與細胞包覆。研究亦證實其包覆細胞培養系統有助軟骨細胞維持其型態與活性。本研究將利用大分子 (25 KDa)、具電中性 (等電6~6.25) 之已被證實具有生物活性含纖維素結合區域之RGD (cellulose-binding domain linked Arg-Gly-Asp, CBD-RGD) 之融合蛋白 ( RGD-Chimeric protein)，結合此褐藻膠培養系統，測試其對大鼠脂肪幹細胞分化為軟骨之影響。研究證實CBD-RGD能專一性結合於細胞表面，經七天培養仍可分別保留20.18±0.73%和8.6±2.17% 蛋白於10mg/g 和 20 mg/g包裹濃度。比較包裹不同濃度之蛋白 (10 and 20 mg/g)，濃度10 mg/g能經由結合膜表面專一受器(integrin α5 and β1-dependent pathway) 顯著提升生長因子 (TGF-3β) 誘導軟骨細胞之分化成效，包含促進早期軟骨分化基因表現 (Sox9, Aggrecan, fibronectin and collagen type II) 與胞外基質 (fibronetin、collagen type II and glycosaminoglycans) ，並同時抑制經由TGF-β3誘導之肥大軟骨細胞基因Collagen type X表現。相反地，濃度20 mg/g則會顯著抑制分化效果。研究發現，CBD-RGD 促進分化程度與Sox9 基因表現之相關性高於其它軟骨基因，且會暫時抑制第一天fibronetin 蛋白和持續性抑制RhoA 活性表現，故實驗結果推測，CBD-RGD促進細胞分化基轉可能與調控RhoA訊息傳遞有關。因此，本實驗成功建立有效便利之CBD-RGD結合褐藻膠三維培養系統，利用蛋白能保持於水膠中 (protein-depot)亦能緩慢釋放，促使軟骨分化過程中包裹之細胞能經由彈性調分化環境有效促進其分化之表現。|
Label of human bone mesenchymal stem cells with CdSe/ZnS quantum dots (QDs) had been demonstrated to impair cell functions and activities. In the present study, QDs delivered by two different routes, Pep-1-labeled QDs (LQ) and PolyFect transfected QDs (TQ), were utilized to assess the effects of delivery mechanisms on various cellular responses of the QDs-internalized human adipose-derived adult stem cells (hADAS). Examination of labeled cells by flow cytometry and laser scanning confocal microscopy showed that LQ had higher fluorescence intensity due to the culster formation and their distribution in cytoplama while TQ were preferentially accumulated at peri-nuclear regions. The fluorescence intensity of the LQ group was still higher than that of the TQ group at 28 days after labeling, though cellular LQ were partitioned after initial cell division. Pep-1 but not PolyFect delivery facilitated QDs to escape from lysosome degradation. Pep-1 delivery of QDs rescued the cells from the negative effects caused by the internalized QDs on cell proliferation and on the expressions of CD29 and CD105 as well as osteogenic and chondrogenic associated lineage markers. The same effect was also observed in the expression of alkaline phosphatase activity, calcium deposition and secretion of chondrogenic matrices (GAG and collagen type II) in micromass culture. These indicated that Pep-1-delivered QDs may serve appropriately to track the hADAS employed in cell therapy/tissue engineering applications. A further study was conducted to reveal the associated mechanism. By comparing Pep-1- and PolyFect-mediated QD internalizations, the connexin 43 (Cx43)-mediated gap junction intercellular communication (GJIC) of hADAS was investigated in monolayer and in three-dimensional (3D) culture (alginate hollow spheres). The latter system offered cells more mobility, which was more similar as in vivo. The results showed that Pep-1-coated QDs, which escaped from the endo-/lysosome degradation, could activate the F-actin assembly and the ERK-dependent phosphorylation of Cx43. The consequence was a reduction in Cx43-mediated GJIC. When the cells were grown in high density 3D alginate hollow spheres instead of in monolayer, the decrease of GJIC caused by the QD internalization was restored. These results indicated that the adaptability in QDs-mediated regulation of GJIC with different delivery coatings depended on the culture systems. The study also suggested that the regulation of gap junction may play a key role in QD cytotoxicity. On the other hand, the role of integrin-binding peptides RGD on chondrogenesis of mesenchymal stem cells is controversial. We revealed the feasibility for flexible modification of RGD by embedding a large molecular weight and slightly charged (isoelectric point, 6-6.25) RGD-chimeric protein (CBD-RGD) with cellulose-binding domain (CBD) in three dimensional (3D) alginate beads to evaluate the chondrogenesis of ADAS. The binding of CBD-RGD with cells and its diffusion from alginate beads were studied on fluorescein isothiocyanate (FITC)-conjugated CBD-RGD. The increases in gene expression (Sox9, Aggrecan, fibronectin and collagen II), accumulation of chondrogenic matrice and decrease of collagen X gene expression during TGF-β3 induction were only observed for those beads containing 10 mg/g CBD-RGD initially, with 20.18±0.73% of that released in a week. The contrary was observed for beads with CBD-RGD 20 mg/g initially and having higher persistence (only 8.6±2.17% relased in a week). The 10 mg/g CBD-RGD-meiated enhancement was demonstrated via the activation of integrin α5 and β1-dependent pathway, and especially related to the upregulation of Sox9 gene and the temporary block of fibronectin expression as well as sustained inhibition of RhoA activity in the early differentiation stage. Thus, we speculated that the dynamic mobility of CBD-RGD may account for the enhanced chondrogenesis. It was concluded that the CBD-RGD-alginate culture system promoted the chondrogenesis of mesenchymal stem cells coordinated with TGF-β3 induction in an RGD dose-dependent manner.
|Appears in Collections:||生命科學系所|
Show full item record
TAIR Related Article
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.