In Vitro Induction of Directional Differentiation of Bone Marrow Mesenchymal Stem Cells towards Chondrocytes_Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

Authors：

TAN Rongbang , SHI Hongcan.

Department of Thoracic Surgery，Medical College of Yangzhou University，Yangzhou 225001，Jiangsu，P. R. China;

Corresponding author：

Keywords：

Mesenchymal stem cell; 　Chondrocyte differentiation; 　Ultrastructure; 　Collagen type Ⅱ

DOI：

10.7507/1007-4848.20130097

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To isolate，culture and expand bone marrow mesenchymal stem cells （MSCs） in vitro，induce MSCs to differentiate directionally towards chondrocytes，and provide experimental basis for clinical application of MSCs and construction of tissue engineering tracheal cartilage. Methods Cultured MSCs were isolated from bone marrow　of Sprague-Dawley rats，purified using adherence separation，and identified by flow cytometry analysis. Transforming growth factor β1 （TGF-β1）and insulin-like growth factor 1 （IGF-1） were used as main induction factors to induce MSCs to differentiate directionally towards chondrocytes. The expression of collagen typeⅡwas evaluated by immunocytochemical staining 21 days after induction. Light microscope and electron microscope were used to observe tiny and ultrastructural changes of the cells before and after induction. Results The expression of collagen typeⅡwas positive by immunocytochemical staining 21 days after induction. MSCs were fusiform before induction under light microscope and electron　microscope. After induction，the cells became larger，polygon，star-shaped or triangular. Transmission electron microscope showed that the cells had abundant organelles，larger nuclei and more nucleoli after induction. Conclusion Abundant　organelles，larger nuclei and more nucleoli are the ultrastructure changes of chondrocytes differentiated from MSCs，indicating that the cells are active in differentiation and metabolism.

Citation： TAN Rongbang,SHI Hongcan.. In Vitro Induction of Directional Differentiation of Bone Marrow Mesenchymal Stem Cells towards Chondrocytes. Chinese Journal of Clinical Thoracic and Cardiovascular Surgery, 2013, 20(3): 325-328. doi: 10.7507/1007-4848.20130097 Copy

1.	Lian QZ， Lye E， Suan Y K， et al. Derivation of clinically compliant　MSCs from CD105＋， CD24- differentiated human ESCs. Stem Cells， 2007， 25 （2）：425-436.
2.	Deng W， Obrocka M， Fischer I， et al. In vitro differentiation of human marrow stromal cells into early progenitors of neural cells by conditions that increase intracellular cyclic AMP. Biochem Biophys Res Commun， 2001， 282 （1）：148-152.
3.	Kopen GC， Prockop DJ， Phinney DG. Marrow stromal cells migrate throughout forebrain and cerebellum， and they differentiate into　astrocytes after injection into neonatal mouse brains. Proc Natl Acad Sci U S A， 1999， 96 （19）：10711-10716.
4.	何凤璞，汪黎明，陈文，等. 大鼠骨髓间充质干细胞体外诱导分化为血管平滑肌样细胞的实验研究. 中国胸心血管外科临床杂志， 2011， 18 （3）：236-239.
5.	Imanishi Y， Saito A， Komoda H， et al. Allogenic mesenchymal stem cell transplantation has a therapeutic effect in acute myocardial infarction in rats. J Mol Cell Cardiol， 2008， 44 （4）：662-671.
6.	Huang NF， Li S. Mesenchymal stem cells for vascular regeneration. Regen Med， 2008， 3 （6）：877-892.
7.	Friendenstein AJ， Petrakovak V. Heterotopic transplants of bone marrow. Analysis of precursor cells for osteogenic and hematotoxic tissues. Transplantation， 1968， 6 （4）：230- 247.
8.	Lin HT， Tang YW， Chen YC， et al. Using human plasma supplem-　ented medium to cultivate human bone marrow-derived mesenchymal stem cell and evaluation of its multiple-lineage potential. Transplant Proc， 2005， 37 （10）：4504-4505.
9.	Bussolati B， Camussi G. Adult stem cells and renal repair. J Nephrol，2006， 19 （6）：706-709.
10.	Attisano L， Wrana JL. Signal transduction by the TGF-beta superfamily. Science， 2002， 296 （5573）：1646-1647.
11.	Kawamura K， Chu CR， Sobajima S， et a1. Adenoviral-mediated transfer of TGF-betal but not IGF-1 induces chondrogenic differentiation of human mesenchymaI stem cells in pellet cultures. Exp　Hemato1， 2005， 33 （8）：865-872.
12.	Matsumoto T， Tsurumoto T， Goldring MB， et al. Differential　effects of IGF-binding proteins， IGFBP-3 and IGFBP-5， on IGF-I action and binding to cell membranes of immortalized human chondrocytes. J Endocrinol， 2000， 166 （1）：29-37.
13.	邓进，彭吾训，王蕾，等. 骨髓间充质干细胞二维培养条件下向软骨细胞诱导分化的实验研究. 中国现代医学杂志， 2008， 18 （16）：2340-2343.
14.	Longobardi L， O'Rear L， Aakula S， et al. Effect of IGF-I in the chondrogenesis of bone marrow mesenchymal stem cells in the presence or absence of TGF-beta signaling. J Bone Miner Res， 2006， 21 （4）：626-636.
15.	Fukumoto T， Sperling JW， Sanyal A， et al. Combined effects of insulin-like growth factor-1 and transforming growth factor-beta1 on periosteal mesenchymal cells during chondrogenesis in vitro.　Osteoarthritis Cartilage， 2003， 11 （1）：55-64.
16.	姜辉，王辉山，汪曾炜，等. 人骨髓间充质干细胞体外分离、培养及鉴定的实验研究. 中国胸心血管外科临床杂志， 2010， 17 （1）：41-45.
17.	刘斌，戎利民，董健文，等. 密度梯度离心与贴壁筛选法体外分离培养胎儿骨髓间充质干细胞. 中国组织工程研究与临床康复， 2009， 13 （40）：7913-7918.
18.	Mcbeath R， Pirone DM， Nelson CM， et al. Cell shape， cytoskeletal　tension， and RhoA regulate stem cell lineage commitment. Dev Cell， 2004， 6 （4）：483-495.

1. 　Lian QZ， Lye E， Suan Y K， et al. Derivation of clinically compliant　MSCs from CD105＋， CD24- differentiated human ESCs. Stem Cells， 2007， 25 （2）：425-436.
2. 　Deng W， Obrocka M， Fischer I， et al. In vitro differentiation of human marrow stromal cells into early progenitors of neural cells by conditions that increase intracellular cyclic AMP. Biochem Biophys Res Commun， 2001， 282 （1）：148-152.
3. 　Kopen GC， Prockop DJ， Phinney DG. Marrow stromal cells migrate throughout forebrain and cerebellum， and they differentiate into　astrocytes after injection into neonatal mouse brains. Proc Natl Acad Sci U S A， 1999， 96 （19）：10711-10716.
4. 　何凤璞，汪黎明，陈文，等. 大鼠骨髓间充质干细胞体外诱导分化为血管平滑肌样细胞的实验研究. 中国胸心血管外科临床杂志， 2011， 18 （3）：236-239.
5. 　Imanishi Y， Saito A， Komoda H， et al. Allogenic mesenchymal stem cell transplantation has a therapeutic effect in acute myocardial infarction in rats. J Mol Cell Cardiol， 2008， 44 （4）：662-671.
6. 　Huang NF， Li S. Mesenchymal stem cells for vascular regeneration. Regen Med， 2008， 3 （6）：877-892.
7. 　Friendenstein AJ， Petrakovak V. Heterotopic transplants of bone marrow. Analysis of precursor cells for osteogenic and hematotoxic tissues. Transplantation， 1968， 6 （4）：230- 247.
8. 　Lin HT， Tang YW， Chen YC， et al. Using human plasma supplem-　ented medium to cultivate human bone marrow-derived mesenchymal stem cell and evaluation of its multiple-lineage potential. Transplant Proc， 2005， 37 （10）：4504-4505.
9. 　Bussolati B， Camussi G. Adult stem cells and renal repair. J Nephrol，2006， 19 （6）：706-709.
10. 　Attisano L， Wrana JL. Signal transduction by the TGF-beta superfamily. Science， 2002， 296 （5573）：1646-1647.
11. 　Kawamura K， Chu CR， Sobajima S， et a1. Adenoviral-mediated transfer of TGF-betal but not IGF-1 induces chondrogenic differentiation of human mesenchymaI stem cells in pellet cultures. Exp　Hemato1， 2005， 33 （8）：865-872.
12. 　Matsumoto T， Tsurumoto T， Goldring MB， et al. Differential　effects of IGF-binding proteins， IGFBP-3 and IGFBP-5， on IGF-I action and binding to cell membranes of immortalized human chondrocytes. J Endocrinol， 2000， 166 （1）：29-37.
13. 　邓进，彭吾训，王蕾，等. 骨髓间充质干细胞二维培养条件下向软骨细胞诱导分化的实验研究. 中国现代医学杂志， 2008， 18 （16）：2340-2343.
14. 　Longobardi L， O'Rear L， Aakula S， et al. Effect of IGF-I in the chondrogenesis of bone marrow mesenchymal stem cells in the presence or absence of TGF-beta signaling. J Bone Miner Res， 2006， 21 （4）：626-636.
15. 　Fukumoto T， Sperling JW， Sanyal A， et al. Combined effects of insulin-like growth factor-1 and transforming growth factor-beta1 on periosteal mesenchymal cells during chondrogenesis in vitro.　Osteoarthritis Cartilage， 2003， 11 （1）：55-64.
16. 　姜辉，王辉山，汪曾炜，等. 人骨髓间充质干细胞体外分离、培养及鉴定的实验研究. 中国胸心血管外科临床杂志， 2010， 17 （1）：41-45.
17. 　刘斌，戎利民，董健文，等. 密度梯度离心与贴壁筛选法体外分离培养胎儿骨髓间充质干细胞. 中国组织工程研究与临床康复， 2009， 13 （40）：7913-7918.
18. 　Mcbeath R， Pirone DM， Nelson CM， et al. Cell shape， cytoskeletal　tension， and RhoA regulate stem cell lineage commitment. Dev Cell， 2004， 6 （4）：483-495.

Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

In Vitro Induction of Directional Differentiation of Bone Marrow Mesenchymal Stem Cells towards Chondrocytes

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content