Matrigel Enhances in vitro Bone Differentiation of Human Marrow-derived Mesenchymal Stem Cells

Document Type: Original Article

Authors

1 Department of Stem Cells and Developmental Biology, Royan Institute for Stem Cell biology and Technology, ACECR, Tehran, Iran

2 Department of Stem Cells and Developmental Biology, Royan Institute for Stem Cell biology and Technology,ACECR, Tehran, Iran

Abstract

Objective(s)
The use of co-culture cells as well as extra cellular matrix are among those strategies that have been
employed to direct mesenchymal stem cell (MSC) bone differentiation in culture. In this regard, there is no study considering the effects of Matrigel on mesenchymal stem cell (MSC) in vitro bone differentiation. This was the subject of the present study.
Materials and Methods
Human passaged-3 MSCs isolated from the marrow aspirates were seeded on either Matrigel or conventional polystyrene plastic surfaces (as control) for 10 days. To compare the cell proliferation in two cultures, the cell numbers were determined during the cultivation period. For bone differentiation, the confluent cultures from either group were provided with osteogenic medium and incubated for 21 days during which the alkaline phosphates (ALP) activity, culture mineralization and the expression of some bone-related genes were quantified and statistically compared.
Results
MTT assay indicated that Matrigel-cultivated cells underwent statistically less proliferation than polystyrenecultivated cells (P<0.05). Regarding the osteogenic differentiation, ALP activity was significantly high in Matrigel versus plastic cultures. Calcium deposition in Matrigel cultures tended to be significantly extensive compared with that of control cultures (2.533±0.017 versus 0.607±0.09 mM). Furthermore, according to the semi-quantitative RT-PCR analysis, compared with polystyrene plastic surface, Matrigel seemed to provide a microenvironment in which human MSC expressed osteocalcin and collagen I genes in a significantly higher level.
Conclusion
Collectively it seems that Matrigel could be considered as an appropriate matrix for MSC osteogenic
differentiation.

Keywords


1.Hay ED. Cell Biology of the Extracellular Matrix. 2nd ed. New York: Plenum Press; 1991.

2.Vukicevic S, Kleinman HK, Luyten FP, Roberts AB, Roche NS, Reddi AH. Identification of multiple active growth factors in basement membrane Matrigel suggests caution in interpretation of cellular activity related to extracellular matrix components. Exp Cell Res 1992; 202:1-8.

3.Fuller GM, Shields D. Cell junction, cell-cell adhesion, and extracellular matrix. In: Fuller GM, Shields D, editors. Molecular Basis of Medical Cell Biology. Connecticut USA: Appleton & Lange, Stamford; 1998. p. 148-166.

4.Streuli CH, Bailey N, Bissell MJ. Control of mammary epithelial differentiation: Basement membrane induces tissue-specific gene expression in the absence of cell-cell interaction and morphological polarity. J Cell Biol 1991; 5:1383-1396.

5.Kleinman HK, Schnaper HW. Basement membrane matrices in tissue development. Am J Respir Cell Mol Biol 1993; 8:238-239. 

6.Oktay K, Karlikaya G, Akman O, Ojakian GK, Oktay M. Interaction of extracellular matrix and activin-A in the initiation of follicle growth in the mouse ovary. Biol Reprod 2000; 63:457-461.

7.Aharoni D, Meiri I, Atzmon R, Vlodavsky I, Amsterdam A. Differential effect of components of the extracellular matrix on differentiation and apoptosis. Curr Biol 1997; 7:43-51.

8.Pullan S, Wilson J, Metcalfe A, Edwards GM, Goberdhan N, Tilly J, et al. Requirement of basement membrane for the suppression of programmed cell death in mammary epithelium. J Cell Sci 1996; 109:631-642.

9.Kleinman HK, Martin GR. Matrigel: basement membrane matrix with biological activity. Semin Cancer Biol 2005; 15:378-386.

10.Vukicevic S, Kleinman HK, Luyten FP, Roberts AB, Roche NS, Reddi AH. Identification of multiple active growth factors in basement membrane Matrigel suggests caution in interpretation of cellular activity related to extracellular matrix components. Exp Cell Res 1992; 202:1-8.

11.Hadley MA, Byers SW, Suarez-Quian CA, Kleinman HK, Dym M. Extracellular matrix regulates sertoli cell differentiation, testicular cord formation and germ cell development. J Cell Biol 1985; 101:1511-1522.

12.Eslaminejad MB, Valojerdi MR, Ashtiani SK. A comparison of polarized and non- polarized human endometrial monolayer culture systems on murine embryo development. J Exp clin Assist Reprod 2005; 2:7.

13.Eslaminejad MB, Valojerdi MR, Ashtiani SK, Yazdi PE. Light and electron microscopic study of epithelial cells from human oviduct and uterus sub-cultured on ECM-Gel. J Reprod Med 2007; 52:503-512.

14.Kubota Y, Kleinman HK, Martin GR, Lawley TJ. Role of laminin and basement membrane in the differentiation of human endothelial cells into capillary-like structure. J Cell Biol 1988; 107: 1589-1598.

15.Yang J, Nagavarapu U,Relloma K, Sjaastad MD, Moss WC, Passaniti A, et al. Telomerized human microvasculature is functional in vivo. Nat Biotechnol 2001; 19:219-224.

16.Hoffman MP, Kibbey MC, Letterio JJ, Kleinman HK. Role of laminin-1 and TGF beta-3 in acinar differentiation of a human submandibular gland cell line (HSG). J Cell Sci 1996; 109:2013-2021.

17.Salasznyk RM, Williams WA, Boskey A, Batorsky A, Plopper GE. Adhesion to vitronectin and collagen I promote osteogenic differentiation of human mesenchymal stem cells. J Biomed Biotechnol 2004; 1:24-34.

18.Klees RF, Salasznyk RM, Kingsley K, Williams WA, Boskey A, Plopper GE. Laminin-5 induces osteogenic gene expression in human mesenchymal stem cells through an ERK-dependent pathway. Mol Biol Cell 2005; 16: 881-890.

19.Bradham DM, Passaniti A, Horton W. Mesenchymal cell chondrogenesis is stimulated by basement membrane matrix and inhibited by age-associated factors. Matrix Biol 1995; 14:561-571.

20.Philp D, Chenb SS, Fitzgeraldb W, Orensteinc J, Margolisb L, Kleinman HK. Complex extracellular matrices promote tissue-specific stem cell differentiation. Stem Cells 2005; 23:288-296.

21.Eslaminejad MB, Nazarian H, Falahi F, Taghiyar L, Daneshzadeh MT. Ex vivo expansion and differentiation of mesenchymal stem cells from goat bone marrow. Iran J Basic Med Sci 2009; 12:70-79.

22.Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284:143-147.

23.Pioletti D, Montjovent MC, Zambelli PY, Applegate L. Bone tissue engineering using fetal cell therapy. Swiss Med Wkly 2002; 136:557-560.

24.Weissman IL. Translating stem and progenitor cell biology to the clinic: barrier and opportunities. Science 2000; 287:1442-1446.

25.Eslaminejed MB, Mirzadeh H, Nickmahzar A, Mohamadi Y, Mivehchi H. Type I collagen gel in seeding medium improves murine mesencymal stem cell loading onto the scaffold, increases their subsequent proliferation and enhances the culture mineralization. J Biomed Mater Res (part B), Appl Biomater 2009; 90B: 659-667.