Human Umbilical Cord Mesenchymal Stem Cells Induce into Chondrocytes in 3D Cultured System

Abstract:

Objective: To investigate a better method of inducing hUC-MSCs into chondrocytes in different culture system in vitro. Method: hUC-MSCs were isolated and cultured by tissue block culture, and the cells surface antigens were identified by flow cytometry, hUC-MSCs were cultured with chondrogenic media and stained with Alcian Blue. The production of matrix was estimated from the determination of hydroxyproline content and Alcian Blue method. Expressions of glycosaminoglycan (GAG), type II collagen and Sox-9 were assayed by real-time fluorescence quantitative PCR. Results: The cultured hUC-MSCs phenotype was CD105+/CD29+/CD44+/ CD31-/CD34-/ CD40-/CD45-/HLA-DR-. hUC-MSCs weakly expressed chondrocyte marker, which strongly expressed GAG and type II collagen after chondrogenic induction, and the cells were incubated in pellet culture with higher expression. Real-time PCR results demonstrated that chondrogenic induction cells were expressed GAG, type II collagen and Sox-9, and the cells were incubated in pellet culture with higher expression. Conclusion: hUC-MSCs incubated in pellet culture is more conducive to differentiate into chondrocytes than those cultured in monolayer culture system.

• Periodical: Advanced Materials Research (Volume 749)
• Pages: 198-205
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.749.198
• Online since: August 2013
• Authors: Li Yu, Jing Liu, Chao Xu, Er Mei Luo, Ming Qiao Tang
• Keywords: Cartilage Tissue Engineering, Chondrocytes, Human Umbilical Cord Mesenchymal Stem Cells, Scaffold

References

[1] Richardson SM, Hoyland JA, Mobasheri R, et al. Mesenchymal stem cells in regenerative medicine: opportunities and challenges forarticular cartilage and intervertebral disc tissue engineering. J Cell Physiol, 2010, 222(1): 23.

DOI: 10.1002/jcp.21915

[2] Vasiliadis HS, Wasiak J. Autologous chondrocyte implantation for full thickness articular cartilage defects of the knee. Cochrane Database Syst Rev, 2010 10: CD003323.

DOI: 10.1002/14651858.cd003323.pub3

[3] Matsumoto T, Okabe T, Ikawa T, et al. Articular cartilage repair with autologous bone marrow mesenchymal cells. J Cell Physiol, 2010, 225(2): 291.

DOI: 10.1002/jcp.22223

[4] Shimomura K, Ando W, Tateishi K, et al. The influence of skeletal maturity on allogenic synovial mesenchymal stem cell-based repair of cartilage in a large animal model. Biomaterials, 2010, 31(31): 8004.

DOI: 10.1016/j.biomaterials.2010.07.017

[5] Majore I, Moretti P, Stahl F, et al. Growth and Differentiation Properties of Mesenchymal Stromal Cell populations derived from Whole Human Umbilical Cord. Stem Cell Rev, 2011, 7(1): 17.

DOI: 10.1007/s12015-010-9165-y

[6] Wegener B, Schrimpf FM, Bergschmidt P, et al. Cartilage regeneration by bone marrow cells-seeded scaffolds. J Biomed Mater Res A, 2010, 95(3): 735.

DOI: 10.1002/jbm.a.32885

[7] Rotter N, Bücheler M, Haisch A, et al. Cartilage tissue engineering using resorbable scaffolds. J Tissue Eng Regen Med, 2007, 1(6): 411.

DOI: 10.1002/term.52

[8] Vinatier C, Mrugala D, Jorgensen C, et al. Cartilage engineering: a crucial combination of cells, biomaterials and biofactors. Trends Biotechnol, 2009, 27(5): 307-314.

DOI: 10.1016/j.tibtech.2009.02.005

[9] Gabay O, Sanchez C, Taboas JM. Update in cartilage bio-engineering. Joint Bone Spine, 2010, 77(4): 283.

DOI: 10.1016/j.jbspin.2010.02.029

[10] Björnsson S. Simultaneous preparation and quantitation of proteoglycans by precipitation with alcian blue. Anal Biochem, 1993, 210(2): 282.

DOI: 10.1006/abio.1993.1197

[11] Tichopad A, Dilger M, Schwarz G, et al. Standardized determination of real-time PCR efficiency from a single reaction set-up. Nucleic Acids Res, 2003, 31(22): 6688.

DOI: 10.1093/nar/gng122

[12] CHEN Lei, YANG Zhiming. Research progress of the seed cells of tissue-engineered cartilage. Chinese Journal of Reparative and Reconstructive Surgery，2003, 17(2): 143.

[13] Wang HS, Hung SC, Peng ST, et al. Mesenchymal Stem Cells in the Wharton's Jelly of the Human Umbilical Cord. Stem Cells, 2004, 22(7): 1330.

DOI: 10.1634/stemcells.2004-0013

[14] Qiao C, Xu W, Zhu W, et al. Human mesenchymal stem cells isolated from the umbilical cord. Cell Biol Int, 2008, 32(1): 8.

[15] Wulf GG, Chapuy B, Trumper L. Mesenchymal stem cells from bone marrow phenotype. Aspects of biology, and clinical perspectives. Med Klin (Munich), 2006, 101(5): 408.

[16] Wernike E, Li Z, Alini M, Grad S, et al. Effect of reduced oxygen tension and long-term mechanical stimulation on chondrocyte-polymer constructs. Cell Tissue Res, 2008, 331(2): 473.

DOI: 10.1007/s00441-007-0500-9

[17] Budde S, Jagodzinski M, Wehmeier M, et al. No effect in combining chondrogenic predifferentiation and mechanical cyclic compression on osteochondral constructs stimulated in a bioreactor. Ann Anat, 2010, 192(4): 237.

DOI: 10.1016/j.aanat.2010.04.001

[18] Furumatsu T, Shukunami C, Amemiya-Kudo M, et al. Scleraxis and E47 cooperatively regulate the Sox9-dependent transcription. Int J Biochem Cell Biol, 2010, 42(1): 148.

DOI: 10.1016/j.biocel.2009.10.003

[19] Topol L, Chen W, Song H, et al. Sox9 inhibits Wnt signaling by promoting beta-catenin phosphorylation in the nucleus. J Biol Chem, 2009, 284(5): 3323.

DOI: 10.1074/jbc.m808048200

[20] Lorda-Diez CI, Montero JA, Martinez-Cue C, et al. Transforming growth factors beta coordinate cartilage and tendon differentiation in the developing limb mesenchyme. J Biol Chem, 2009, 284(43): 29988.

DOI: 10.1074/jbc.m109.014811