Differentiation of Bone Marrow Mesenchymal Stem Cells into Chondrocytes after Short Term Culture in Alkaline Medium
-
Farshad Homayouni Moghadam
,
Tahereh Tayebi
,
Maryam Dehghan
,
Gilda Eslami
,
Hamid Nadri
,
Alireza Moradi
,
Hassanali Vahedian-Ardakani
,
Kazem Barzegar
Abstract
Background: Bone marrow mesenchymal stem cells (MSCs) are one of the undifferentiated multipotential cell sources of human body.
Methods: MSCs have the capacity to form a variety of cell types, especially chondrocytes and osteocytes. Learning about responses of MSCs to external milieu and chemical factors such as pH could recommend new approaches for preparation of suitable scaffolds for bone and cartilage tissue engineering. In present study, the effect of alkaline medium on chondrogenic and osteogenic differentiation of rat MSCs was evaluated.MSCs were harvested from bone marrow of animals and then the response of passage1 and 2 of MSCs (P1 MSCs & P2 MSCs) to the culture in alkaline medium (pH: 8) was evaluated. Cytochemical and immunocytochemical staining were performed to distinguish chondrocytes and osteocytes. Real-time PCR was performed to evaluate the type II collagen and osteopontin mRNA levels.
Results: Staining for type II collagen, a chondrocytic specific marker, revealed that after one-week culture in alkaline medium, a considerable amount of P1 MSCs had shown chondrocytic morphology. By prolonging the culture period up to 4 weeks, osteogenic cells with expanded matrix and mineralized areas around them were appeared. Results of real-time PCR showed that P1 MSCs after one week culture in alkaline medium expressed highest rate of type II collagen and osteopontin mRNA among all groups.
Conclusion: This study demonstrated that alkaline medium is a potent chondrogenic differentiation inducer for MSCs in their first passage.
Giuliani N, Lisignoli G, Magnani M, Racano C, Bolzoni M, Dalla Palma B, Spolzino A, Manferdini C, Abati C, Toscani D, Facchini A, Aversa F. New insights into osteogenic and chondrogenic differentiation of human bone marrow mesenchymal stem cells and their potential clinical applications for bone regeneration in pediatric orthopaedics. Stem Cells Int. 2013; 2013: 312501.
Mauck RL, Yuan X, Tuan RS. Chondrogenic differentiation and functional maturation of bovine mesenchymal stem cells in long-term agarose culture. Osteoarthritis and cartilage / OARS, Osteoarthritis Research Society. 2006; 14: 179-89.
Veronesi F, Giavaresi G, Tschon M, Borsari V, Nicoli Aldini N, Fini M. Clinical use of bone marrow, bone marrow concentrate, and expanded bone marrow mesenchymal stem cells in cartilage disease. Stem Cells Dev. 2013; 22: 181-92.
Zheng YH, Xiong W, Su K, Kuang SJ, Zhang ZG. Multilineage differentiation of human bone marrow mesenchymal stem cells in vitro and in vivo. Exp Ther Med. 2013; 5: 1576-80.
Zhou J, Xu C, Wu G, Cao X, Zhang L, Zhai Z, Zheng Z, Chen X, Wang Y. In vitro generation of osteochondral differentiation of human marrow mesenchymal stem cells in novel collagen-hydroxyapatite layered scaffolds. Acta Biomater. 2011; 7: 3999-4006.
Barry F, Boynton RE, Liu B, Murphy JM. Chondrogenic differentiation of mesenchymal stem cells from bone marrow: differentiation-dependent gene expression of matrix components. Experimental cell research. 2001; 268: 189-200.
Giannotti S, Bottai V, Ghilardi M, Dell'osso G, Fazzi R, Trombi L, Petrini M, Guido G. Treatment of pseudoarthrosis of the upper limb using expanded mesenchymal stem cells: a pilot study. Eur Rev Med Pharmacol Sci. 2013; 17: 224-7.
Manning CN, Schwartz AG, Liu W, Xie J, Havlioglu N, Sakiyama-Elbert SE, Silva MJ, Xia Y, Gelberman RH, Thomopoulos S. Controlled delivery of mesenchymal stem cells and growth factors using a nanofiber scaffold for tendon repair. Acta Biomater. 2013; 9: 6905-14.
Moroz A, Bittencourt RA, Almeida RP, Felisbino SL, Deffune E. Platelet lysate 3D scaffold supports mesenchymal stem cell chondrogenesis: an improved approach in cartilage tissue engineering. Platelets. 2013; 24: 219-25.
Indrawattana N, Chen G, Tadokoro M, Shann LH, Ohgushi H, Tateishi T, Tanaka J, Bunyaratvej A. Growth factor combination for chondrogenic induction from human mesenchymal stem cell. Biochemical and biophysical research communications. 2004; 320: 914-9.
Uebersax L, Merkle HP, Meinel L. Insulin-like growth factor I releasing silk fibroin scaffolds induce chondrogenic differentiation of human mesenchymal stem cells. Journal of controlled release : official journal of the Controlled Release Society. 2008; 127: 12-21.
Ichinose S, Tagami M, Muneta T, Mukohyama H, Sekiya I. Comparative sequential morphological analyses during in vitro chondrogenesis and osteogenesis of mesenchymal stem cells embedded in collagen gels. Med Mol Morphol. 2013; 46: 24-33.
Grässel S, Stöckl S, Jenei-Lanzl Z. Isolation, culture, and osteogenic/chondrogenic differentiation of bone marrow-derived mesenchymal stem cells. Methods Mol Biol. 2012; 879: 203-67.
Zheng YH, Su K, Jian YT, Kuang SJ, Zhang ZG. Basic fibroblast growth factor enhances osteogenic and chondrogenic differentiation of human bone marrow mesenchymal stem cells in coral scaffold constructs. J Tissue Eng Regen Med. 2011; 5: 540-50.
Centola M, Tonnarelli B, Schären S, Glaser N, Barbero A, Martin I. Priming 3D Cultures of Human Mesenchymal Stromal Cells Toward Cartilage Formation Via Developmental Pathways. Stem Cells Dev. 2013.
Augello A, De Bari C. The regulation of differentiation in mesenchymal stem cells. Hum Gene Ther. 2010; 21: 1226-38.
Ruedel A, Hofmeister S, Bosserhoff AK. Development of a model system to analyze chondrogenic differentiation of mesenchymal stem cells. Int J Clin Exp Pathol. 2013; 6: 3042-8.
Bian L, Zhai DY, Mauck RL, Burdick JA. Coculture of human mesenchymal stem cells and articular chondrocytes reduces hypertrophy and enhances functional properties of engineered cartilage. Tissue Eng Part A. 2011; 17: 1137-45.
Buxton AN, Bahney CS, Yoo JU, Johnstone B. Temporal exposure to chondrogenic factors modulates human mesenchymal stem cell chondrogenesis in hydrogels. Tissue Eng Part A. 2011; 17: 371-80.
Farrell MJ, Fisher MB, Huang AH, Shin JI, Farrell KM, Mauck RL. Functional properties of bone marrow-derived MSC-based engineered cartilage are unstable with very long-term in vitro culture. J Biomech. 2013.
Sheehy EJ, Buckley CT, Kelly DJ. Oxygen tension regulates the osteogenic, chondrogenic and endochondral phenotype of bone marrow derived mesenchymal stem cells. Biochem Biophys Res Commun. 2012; 417: 305-10.
Kohn DH, Sarmadi M, Helman JI, Krebsbach PH. Effects of pH on human bone marrow stromal cells in vitro: implications for tissue engineering of bone. Journal of biomedical materials research. 2002; 60: 292-9.
Wu MH, Urban JP, Cui ZF, Cui Z, Xu X. Effect of extracellular ph on matrix synthesis by chondrocytes in 3D agarose gel. Biotechnology progress. 2007; 23: 430-4.
Leem YH NT, Kim JH, Lee KS, Lee DH, Yun J, and Chang JS. The Effects of Extracellular pH on Proliferation and Differentiation of human Bone Marrow Stem Cells. Korean Journal of Bone Metabolism. 2012; 19(1): 12.
Ye R, Hao J, Song J, Zhao Z, Fang S, Wang Y, Li J. Microenvironment is Involved in Cellular Response to Hydrostatic Pressures During Chondrogenesis of Mesenchymal Stem Cells. J Cell Biochem. 2013.
Lennon DP, Caplan AI. Isolation of rat marrow-derived mesenchymal stem cells. Exp Hematol. 2006; 34: 1606-7.
Hegert C, Kramer J, Hargus G, Müller J, Guan K, Wobus AM, Müller PK, Rohwedel J. Differentiation plasticity of chondrocytes derived from mouse embryonic stem cells. J Cell Sci. 2002; 115: 4617-28.
Yumoto K, Ishijima M, Rittling SR, Tsuji K, Tsuchiya Y, Kon S, Nifuji A, Uede T, Denhardt DT, Noda M. Osteopontin deficiency protects joints against destruction in anti-type II collagen antibody-induced arthritis in mice. Proc Natl Acad Sci U S A. 2002; 99: 4556-61.
Bushinsky DA. Metabolic alkalosis decreases bone calcium efflux by suppressing osteoclasts and stimulating osteoblasts. The American journal of physiology. 1996; 271: F216-22.
Frick KK, Jiang L, Bushinsky DA. Acute metabolic acidosis inhibits the induction of osteoblastic egr-1 and type 1 collagen. The American journal of physiology. 1997; 272: C1450-6.
Frick KK, Bushinsky DA. Chronic metabolic acidosis reversibly inhibits extracellular matrix gene expression in mouse osteoblasts. The American journal of physiology. 1998; 275: F840-7.
Kaysinger KK, Ramp WK. Extracellular pH modulates the activity of cultured human osteoblasts. Journal of cellular biochemistry. 1998; 68: 83-9.
Krieger NS, Sessler NE, Bushinsky DA. Acidosis inhibits osteoblastic and stimulates osteoclastic activity in vitro. The American journal of physiology. 1992; 262: F442-8.
Lennon DP, Schluchter MD, Caplan AI. The effect of extended first passage culture on the proliferation and differentiation of human marrow-derived mesenchymal stem cells. Stem cells translational medicine. 2012; 1: 279-88.
Sila-Asna M, Bunyaratvej A, Maeda S, Kitaguchi H, Bunyaratavej N. Osteoblast differentiation and bone formation gene expression in strontium-inducing bone marrow mesenchymal stem cell. The Kobe journal of medical sciences. 2007; 53: 25-35.
Varshney RR, Zhou R, Hao J, Yeo SS, Chooi WH, Fan J, Wang DA. Chondrogenesis of synovium-derived mesenchymal stem cells in gene-transferred co-culture system. Biomaterials. 2010; 31: 6876-91.
Solchaga LA, Penick KJ, Welter JF. Chondrogenic differentiation of bone marrow-derived mesenchymal stem cells: tips and tricks. Methods Mol Biol. 2011; 698: 253-78.
Freeman FE, Haugh MG, McNamara LM. Investigation of the optimal timing for chondrogenic priming of MSCs to enhance osteogenic differentiation in vitro as a bone tissue engineering strategy. J Tissue Eng Regen Med. 2013.
Zhu S, Zhang B, Man C, Ma Y, Liu X, Hu J. Combined Effects of Connective Tissue Growth Factor-modified Bone Marrow-Derived Mesenchymal Stem Cells and NaOH-treated PLGA Scaffolds on Repair of Articular Cartilage Defect in Rabbits. Cell Transplant. 2013.
| Files | ||
| Issue | Vol 8, No 4 (2014) | |
| Section | Articles | |
| Keywords | ||
| Alkaline medium Chondrocyte Mesenchymal stem cell Osteoblast | ||
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