title: Covalent immobilization of stem cell factor and stromal derived factor 1α for in vitro culture of hematopoietic progenitor cells.
abstract: Hematopoietic stem cells (HSCs) are currently utilized in the treatment of blood diseases, but widespread application of HSC therapeutics has been hindered by the limited availability of HSCs. With a better understanding of the HSC microenvironment and the ability to precisely recapitulate its components, we may be able to gain control of HSC behavior. In this work we developed a novel, biomimetic PEG hydrogel material as a substrate for this purpose and tested its potential with an anchorage-independent hematopoietic cell line, 32D clone 3 cells. We immobilized a fibronectin-derived adhesive peptide sequence, RGDS; a cytokine critical in HSC self-renewal, stem cell factor (SCF); and a chemokine important in HSC homing and lodging, stromal derived factor 1α (SDF1α), onto the surfaces of poly(ethylene glycol) (PEG) hydrogels. To evaluate the system's capabilities, we observed the effects of the biomolecules on 32D cell adhesion and morphology. We demonstrated that the incorporation of RGDS onto the surfaces promotes 32D cell adhesion in a dose-dependent fashion. We also observed an additive response in adhesion on surfaces with RGDS in combination with either SCF or SDF1α. In addition, the average cell area increased and circularity decreased on gel surfaces containing immobilized SCF or SDF1α, indicating enhanced cell spreading. By recapitulating aspects of the HSC microenvironment using a PEG hydrogel scaffold, we have shown the ability to control the adhesion and spreading of the 32D cells and demonstrated the potential of the system for the culture of primary hematopoietic cell populations.
authors: Department of Bioengineering, Rice University, MS-142, 6100 Main Street, Houston, TX 77005-1892, USA.
authors: Maude L
authors: ML
authors: Cuchiara
authors: Kelsey L
authors: KL
authors: Horter
authors: Omar A
authors: OA
authors: Banda
authors: Jennifer L
authors: JL
authors: West
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000494:pharmacology; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D048430:Cell Shape / Q000187:drug effects; D002478:Cells, Cultured; D054377:Chemokine CXCL12 / Q000494:pharmacology*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D055767:Immobilized Proteins / Q000494:pharmacology*; D051379:Mice; D055616:Microtechnology; D009842:Oligopeptides / Q000494:pharmacology; D010969:Plastics / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D019089:Stem Cell Factor / Q000494:pharmacology*; D013499:Surface Properties
chemical_list: D001672:Biocompatible Materials; D054377:Chemokine CXCL12; D055767:Immobilized Proteins; D009842:Oligopeptides; D010969:Plastics; D019089:Stem Cell Factor; C437167:poly(ethylene glycol)diacrylate; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D011092:Polyethylene Glycols; C046617:arginyl-glycyl-aspartyl-serine
keywords: Hematopoietic cell; Hydrogel; Poly(ethylene glycol); Stem cell factor; Stromal derived factor 1α
references: J Clin Invest. 1992 Aug;90(2):358-67
references: 1379610
references: Acta Biomater. 2011 Jan;7(1):133-43
references: 20801242
references: Exp Hematol. 2007 May;35(5):771-81
references: 17577926
references: Langmuir. 2007 Oct 9;23(21):10635-43
references: 17803326
references: Biotechnol Bioeng. 2006 Apr 20;93(6):1060-8
references: 16444742
references: J Biomed Mater Res A. 2006 Sep 15;78(4):781-91
references: 16739181
references: J Exp Med. 1996 Sep 1;184(3):1101-9
references: 9064327
references: Cell Tissue Res. 2010 Jan;339(1):121-30
references: 19588168
references: Biomaterials. 2008 Jul;29(20):2962-8
references: 18433863
references: Biomaterials. 2006 Apr;27(12):2519-24
references: 16375965
references: J Leukoc Biol. 2002 Mar;71(3):425-32
references: 11867680
references: Exp Hematol. 2001 Mar;29(3):345-55
references: 11274763
references: Blood. 1998 Feb 15;91(4):1101-34
references: 9454740
references: Am J Physiol Cell Physiol. 2005 Jan;288(1):C30-8
references: 15371257
references: Exp Hematol. 2001 Dec;29(12):1456-64
references: 11750105
references: Blood. 1995 Feb 1;85(3):641-9
references: 7530502
references: J Clin Invest. 1992 Jul;90(1):251-5
references: 1378855
references: Nat Biotechnol. 2010 Oct;28(10):1123-8
references: 20890282
references: J Leukoc Biol. 1995 Jul;58(1):14-22
references: 7542304
references: Biomacromolecules. 2007 Jan;8(1):42-9
references: 17206786
references: J Clin Invest. 1996 Jul 15;98(2):521-8
references: 8755665
references: Biomaterials. 2010 Jul;31(21):5491-7
references: 20447685
references: N Engl J Med. 1959 Apr 2;260(14):675-83
references: 13644566
references: Blood. 1993 Jan 15;81(2):344-51
references: 7678511
references: Biol Proced Online. 2009 May 15;11:32-51
references: 19495910
references: J Biol Chem. 1993 Jul 5;268(19):14189-201
references: 7686152
references: Blood. 2000 Jun 1;95(11):3289-96
references: 10828007
references: Biomaterials. 2000 Dec;21(23):2405-12
references: 11055288
references: Biomacromolecules. 2011 Jul 11;12(7):2715-22
references: 21639150
references: Blood. 1990 Dec 1;76(11):2311-20
references: 2147863
references: Leukemia. 1998 Sep;12(9):1375-82
references: 9737685
references: Integr Biol (Camb). 2009 Jan;1(1):59-69
references: 20023792
references: J Clin Invest. 1999 Nov;104(9):1199-211
references: 10545519
references: FASEB J. 2007 Mar;21(3):790-801
references: 17197384
references: Langmuir. 2004 Jun 22;20(13):5153-6
references: 15986641
references: Biomaterials. 2012 Jun;33(18):4460-8
references: 22444641
references: Biomaterials. 2008 May;29(13):2096-103
references: 18289666
references: Adv Drug Deliv Rev. 2002 Jun 17;54(4):459-76
references: 12052709
references: Langmuir. 2009 Aug 4;25(15):8774-9
references: 19361170
references: Biomaterials. 2011 Jan;32(2):321-9
references: 20952054
references: J Clin Invest. 1959 Oct;38:1709-16
references: 13837954
references: J Immunol. 1994 Jan 1;152(1):213-9
references: 7504710
references: Blood. 1992 Oct 1;80(7):1679-84
references: 1382698
references: J Clin Invest. 1991 Jan;87(1):6-11
references: 1824634
references: J Exp Med. 1991 Sep 1;174(3):693-703
references: 1875168
references: J Cell Sci. 2012 Aug 15;125(Pt 16):3765-75
references: 22553208
references: Biomaterials. 2006 May;27(13):2723-32
references: 16376984
references: Blood. 1989 Jan;73(1):111-6
references: 2910353
references: J Leukoc Biol. 2002 Aug;72(2):353-62
references: 12149427
references: Biomaterials. 1998 Jul;19(14):1287-94
references: 9720892
references: EMBO J. 2003 May 15;22(10):2324-33
references: 12743027
references: Biomaterials. 2009 Sep;30(25):4047-52
references: 19481255
references: Science. 1999 Feb 5;283(5403):845-8
references: 9933168
references: Langmuir. 2009 Mar 3;25(5):2994-3002
references: 19437769
references: Cell. 1990 Oct 5;63(1):235-43
references: 1698558
references: J Biol Chem. 1994 Dec 16;269(50):31991-8
references: 7527401
references: Biomaterials. 2009 Feb;30(5):711-20
references: 19010529
references: Cell Tissue Kinet. 1977 Mar;10(2):121-6
references: 856473
references: Biomaterials. 2001 Mar;22(5):405-17
references: 11214751
references: Macromol Rapid Commun. 2012 Sep 14;33(17):1432-8
references: 22815039
references: BioDrugs. 2008;22(5):315-29
references: 18778113
references: J Immunol. 1994 Oct 15;153(8):3717-23
references: 7523504
references: Integr Biol (Camb). 2009 Jun;1(5-6):427-34
references: 20023748
references: Biomaterials. 2005 Jun;26(16):3227-34
references: 15603817
references: Biomaterials. 2006 Dec;27(36):6043-51
references: 16854459
references: Haematologica. 2009 Nov;94(11):1477-81
references: 19880773
references: Blood. 1995 Jan 1;85(1):159-67
references: 7528564
references: Tissue Eng. 2004 Nov-Dec;10(11-12):1775-86
references: 15684686
references: Biomaterials. 2002 Nov;23(22):4315-23
references: 12219821
references: Blood. 1994 Jan 1;83(1):137-45
references: 7506076
references: Cancer Res. 1965 Oct;25(9):1525-31
references: 5323965
references: Proc Natl Acad Sci U S A. 1999 Mar 2;96(5):2087-92
references: 10051599
references: Blood. 2003 Jun 15;101(12):4725-32
references: 12560232
references: J Biomed Mater Res. 2002 Apr;60(1):86-93
references: 11835163
references: J Biomed Mater Res A. 2004 Mar 1;68(3):458-72
references: 14762925
references: Macromol Biosci. 2011 Jun 14;11(6):739-47
references: 21341371
references: J Exp Med. 1995 May 1;181(5):1805-15
references: 7536795
references: Blood. 1990 Feb 15;75(4):865-73
references: 2137353
references: J Biomed Mater Res. 1998 Feb;39(2):266-76
references: 9457557
references: J Biomater Sci Polym Ed. 2009;20(12):1763-79
references: 19723440
references: Blood. 1997 Sep 1;90(5):1858-66
references: 9292518
references: J Exp Med. 1997 Jan 6;185(1):111-20
references: 8996247
references: Biomaterials. 2003 Nov;24(24):4385-415
references: 12922151
references: J Biomed Mater Res B Appl Biomater. 2010 Jan;92(1):32-9
references: 19637375
references: Exp Hematol. 1999 Mar;27(3):496-504
references: 10089912
references: J Am Chem Soc. 2004 Nov 24;126(46):15223-30
references: 15548019
references: Proc Natl Acad Sci U S A. 1985 Jan;82(2):440-4
references: 3855563
references: Biomaterials. 2012 Apr;33(11):3107-18
references: 22269650
references: Blood. 1998 May 1;91(9):3263-72
references: 9558382
references: Immunity. 2006 Dec;25(6):977-88
references: 17174120
references: Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7350-4
references: 1380155