, Volume 66, Issue 4, pp 575–584 | Cite as

Cardiomyocyte-like cells differentiation from non β-catenin expression mesenchymal stem cells

  • Qing GaoEmail author
  • Xiantong Hu
  • Xijuan Jiang
  • Maojuan Guo
  • Hong Ji
  • Yijing Wang
  • Yingchang Fan
Original Research


Recent studies have shown that block wnt/β-catenin signaling pathway is integrant for cardiomyocytes differentiation from bone marrow mesenchymal stem cells (MSCs). By transducing the MSCs with lentivirus which contain β-catenin interference RNA, we screened out the non β-catenin expression clone. In the establishment of knockdown β-catenin in MSCs, we investigated the role of 5-azacytidine (5-aza), salvianolic acid B (salB), and cardiomyocytes lysis medium (CLM) in inducing MSCs to differentiate into cardiomyocyte-like cells. A method for culturing MSCs and cardiomyocytes was established. Purified MSCs were investigated by flow cytometry. The MSCs were positive for CD90 and CD29, but negative for CD34 and CD45. Meanwhile, the cardiomyocytes contracted spontaneously after 24 h of seeding into the plates. The fourth-passage non-β-catenin expression MSCs were divided into eight groups: control group, 5-aza, salB, CLM, 5-aza + salB, 5-aza + CLM, salB + CLM, and 5-aza + salB + CLM. The gene and protein expression of cTnT, α-actin, β-myosin, β-catenin, and GSK-3β were detected by quantitative real-time PCR and Western blotting. Our results showed that cTnT expression in 5-aza + salB + CLM group was ninefold higher than in the control group in the non-β-catenin MSCs model, implying that cardiomyocytes differentiation from MSCs is an extremely complicated process and it is necessary to consider the internal and external environmental conditions, such as suitable pharmaceutical inducers, cardiomyocytes microenvironments, inhibition of the negative signaling pathway and so on.


Mesenchymal stem cells Cardiomyocytes Differentiation Wnt/β-catenin signaling 



This project was supported by the National Natural Science Foundation of China (No. 81173413,, and the Specialized Research Fund for the Doctoral Program of Higher Education of China (Nos. 20101210110004 and 20101210120008,, and Tianjin Research program of Application Foundation and Advanced Technology (No. 09JCYBJC12200, The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Supplementary material

Supplementary material 1 (AVI 2538 kb)


  1. Alfaro MP, Pagni M, Vincent A, Atkinson J, Hill MF, Cates J, Davidson JM, Rottman J, Lee E, Young PP (2008) The Wnt modulator sFRP2 enhances mesenchymal stem cell engraftment, granulation tissue formation and myocardial repair. Proc Natl Acad Sci USA 105:18366–18371CrossRefGoogle Scholar
  2. Chen T, Liu WB, Chao XD, Zhang L, Qu Y, Huo JL, Zhou F (2011) Salvianolic acid B attenuates brain damage and inflammation after traumatic brain injury in mice. Brain Res Bull 84:163–168CrossRefGoogle Scholar
  3. Cho J, Rameshwar P, Sadoshima J (2009) Distinct roles of glycogen synthase kinase (GSK)-3α and GSK-3β in mediating cardiomyocyte differentiation in murine bone marrow-derived mesenchymal stem cells. J Biol Chem 284:36647–36658CrossRefGoogle Scholar
  4. Cho J, Zhai P, Maejima Y, Sadoshima J (2011) Myocardial injection with GSK-3β—overexpressing bone marrow-derived mesenchymal stem cells attenuates cardiac dysfunction after myocardial infarction. Circ Res 108:478–489CrossRefGoogle Scholar
  5. Dell’Ovo V, Bandi E, Coslovich T, Florio C, Sciancalepore M, Decorti G, Sosa S, Lorenzon P, Yasumoto T, Tubaro A (2008) In vitro effects of yessotoxin or a primary culture of rat cardiomyocytes. Toxicol Sci 106:392–399CrossRefGoogle Scholar
  6. Gwak J, Cho M, Gong SJ, Won J, Kim DE, Kim EY, Lee SS, Kim M, Kim TK, Shin JG, Oh S (2006) Protein-kinase-C-mediated β-catenin phosphorylation negatively regulates the wnt/β-catenin pathway. J Cell Sci 119:4702–4709CrossRefGoogle Scholar
  7. Hamidouche Z, Haÿ E, Vaudin P, Charbord P, Schüle R, Marie PJ, Fromigué O (2008) FHL2 mediates dexamethasone-induced mesenchymal cell differentiation into osteoblasts by activating Wnt/beta-catenin signaling-dependent Runx2 expression. FASEB J 22:3813–3822CrossRefGoogle Scholar
  8. Isacchi B, Fabbri V, Galeotti N, Bergonzi MC, Karioti A, Ghelardini C, Vannucchi MG, Bilia AR (2011) Salvianolic acid B and its liposomal formulations: anti-hyperalgesic activity in the treatment of neuropathic pain. Eur J Pharm Sci 44:552–558CrossRefGoogle Scholar
  9. Kilian KA, Bugarija B, Lahn BT, Mrksich M (2010) Geometric cues for directing the differentiation of mesenchymal stem cells. Proc Natl Acad Sci USA 107:4872–4877CrossRefGoogle Scholar
  10. Kim DH, Park SJ, Kim JM, Jeon SJ, Kim DH, Cho YW, Son KH, Lee HJ, Moon JH, Cheong JH, Ko KH, Ryu JH (2011) Cognitive dysfunctions induced by a cholinergic blockade and Aβ 25–35 peptide are attenuated by salvianolic acid B. Neuropharmacology 61:1432–1440CrossRefGoogle Scholar
  11. Liu Y, Song J, Liu W, Wan Y, Chen X, Hu C (2003) Growth and differentiation of rat bone marrow stromal cells: Does 5-azacytidine trigger their cardiomyogenic differentiation? Cardiovasc Res 58:460–468CrossRefGoogle Scholar
  12. Mercola M, Ruiz-Lozano P, Schneider MD (2011) Cardiac muscle regeneration: lessons from development. Genes Dev 25:299–309CrossRefGoogle Scholar
  13. Qiang YW, Hu B, Chen Y, Zhong Y, Shi B, Barlogie B, Shaughnessy JD Jr (2009) Bortezomib induces osteoblast differentiation via Wnt-independent activation of β-catenin/TCF signaling. Blood 113:4319–4330CrossRefGoogle Scholar
  14. Schittini AV, Celedon PF, Stimamiglio MA, Krieger M, Hansen P, da Costa FD, Goldenberg S, Dallagiovanna B, Correa A (2010) Human cardiac explant-conditioned medium: soluble factors and cardiomyogenic effect on mesenchymal stem cells. Exp Biol Med 235:1015–1024CrossRefGoogle Scholar
  15. Schuleri KH, Feigenbaum GS, Centola M, Weiss ES, Zimmet JM, Turney J, Kellner J, Zviman MM, Hatzistergos KE, Detrick B, Conte JV, McNiece I, Steenbergen C, Lardo AC, Hare JM (2009) Autologous mesenchymal stem cells produce reverse remodelling in chronic ischaemic cardiomyopathy. Eur Heart J 30:2722–2732CrossRefGoogle Scholar
  16. Schuleri KH, Feigenbaum GS, Centola M, Weiss ES (2010) Combining neuropeptide Y and mesenchymal stem cells reverses remodeling after myocardial infarction. Am J Physiol Heart Circ Physiol 298:275–286CrossRefGoogle Scholar
  17. Song H, Hwang HJ, Chang W, Song BW, Cha MJ, Kim IK, Lim S, Choi EJ, Ham O, Lee CY, Park JH, Lee SY, Choi E, Lee C, Lee M, Lee MH, Kim SH, Jang Y, Hwang KC (2011) Cardiomyocytes from phorbol myristate acetate activated mesenchymal stem cells restore electromechanical function in infarcted rat hearts. Proc Natl Acad Sci USA 108:296–301CrossRefGoogle Scholar
  18. Ueno S, Weidinger G, Osugi T, Kohn AD, Golob JL, Pabon L, Reinecke H, Moon RT, Murry CE (2007) Biphasic role for Wnt/β-catenin signaling in cardiac specification in zebrafish and embryonic stem cells. Proc Natl Acad Sci USA 104:9685–9690CrossRefGoogle Scholar
  19. Wakitani S, Saito T, Caplan AI (1995) Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-azacytidine. Muscle Nerve 18:1417–1426CrossRefGoogle Scholar
  20. Wei F, Wang T, Liu J, Du Y, Ma A (2011) The subpopulation of mesenchymal stem cells that differentiate toward cardiomyocytes is cardiac progenitor cells. Exp Cell Res 317:2661–2670CrossRefGoogle Scholar
  21. Xing Y, Lv A, Wang L, Yan X (2012) The combination of angiotensin II and 5-azacytidine promotes cardiomyocyte differentiation of rat bone marrow mesenchymal stem cells. Mol Cell Biochem 360:279–287CrossRefGoogle Scholar
  22. Xu L, Deng Y, Feng L, Li D, Chen X, Ma C, Liu X, Yin J, Yang M, Teng F, Wu W, Guan S, Jiang B, Guo D (2011) Cardio-protection of salvianolic acid B through inhibition of apoptosis network. PLoS One 6:e24036CrossRefGoogle Scholar
  23. Zelarayán LC, Noack C, Sekkali B, Kmecova J, Gehrke C, Renger A, Zafiriou MP, van der Nagel R, Dietz R, de Windt LJ, Balligand JL, Bergmann MW (2008) β-Catenin downregulation attenuates ischemic cardiac remodeling through enhanced resident precursor cell differentiation. Proc Natl Acad Sci USA 105:19762–19767CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Qing Gao
    • 1
    Email author
  • Xiantong Hu
    • 1
  • Xijuan Jiang
    • 1
  • Maojuan Guo
    • 1
  • Hong Ji
    • 1
  • Yijing Wang
    • 1
  • Yingchang Fan
    • 1
  1. 1.Key Laboratory of Pathology of State Administration of Traditional Chinese Medicine, School of Traditional Chinese MedicineTianjin University of Traditional Chinese MedicineTianjinChina

Personalised recommendations