Cell and Tissue Banking

, Volume 15, Issue 3, pp 439–450 | Cite as

Isolation and characterization of mesenchymal stem cells derived from fetal bovine liver

  • Taofeng Lu
  • Pengfei Hu
  • Xiaohua Su
  • Changli Li
  • Yuehui Ma
  • Weijun Guan
Original Paper


Bovine liver-derived mesenchymal stem cells (bLMSCs) were isolated from the liver tissue of 4–6 months old fetal calf, and then characterized by immunofluorescence and RT-PCR. We found that primary bLMSCs could be subcultured to 44 passages, the total culture time in vitro was 192 days. The results of surface antigen detection showed that bBMSCs expressed CD29, CD44, CD73, CD90, CD106 and CD166 but not expressed endothelial cells and hematopoietic cells specific marker CD34, CD45 and BLA-DR. The results of growth kinetics, colony-forming cell assay and cell cycle analysis indicated that the fetal bovine LMSCs had good proliferation ability in vitro. The cells from passages 7 were successfully induced to differentiate into osteoblasts, adipocytes and chondrocytes. The results indicate the potential for multi-lineage differentiation of bLMSCs that may represent an ideal candidate for cellular transplantation therapy.


Bovine LMSCs Proliferation Multi-lineage differentiation 



This work was supported by the project National Infrastructure of Animal Germplasm Resources (2013) and Chinese Postdoctoral Science Foundation (2011M500460).


  1. Aquino JB, Bolontrade MF, Garcia MG, Podhajcer OL, Mazzolini G (2010) Mesenchymal stem cells as therapeutic tools and gene carriers in liver fibrosis and hepatocellular carcinoma. Gene Ther 17(6):692–708. doi: 10.1038/gt.2010.10 PubMedCrossRefGoogle Scholar
  2. Arthur A, Zannettino A, Gronthos S (2009) The therapeutic applications of multipotential mesenchymal/stromal stem cells in skeletal tissue repair. J Cell Physiol 218(2):237–245. doi: 10.1002/jcp.21592 PubMedCrossRefGoogle Scholar
  3. Barry F, Boynton RE, Liu B, Murphy JM (2001) Chondrogenic differentiation of mesenchymal stem cells from bone marrow: differentiation-dependent gene expression of matrix components. Exp Cell Res 268(2):189–200. doi: 10.1006/excr 2001.5278PubMedCrossRefGoogle Scholar
  4. Bianco P, Robey PG (2001) Stem cells in tissue engineering. Nature 414(6859):118–121. doi: 10.1038/35102181 PubMedCrossRefGoogle Scholar
  5. Black IB, Woodbury D (2001) Adult rat and human bone marrow stromal stem cells differentiate into neurons. Blood Cells Mol Dis 27(3):632–636. doi: 10.1006/bcmd.2001.0423 PubMedCrossRefGoogle Scholar
  6. Bradstreet RW, Toledo-Pereyra LH (2013) Role of mesenchymal stem cells in partial liver transplantation. J Surg Res. doi: 10.1016/j.jss.2013.04.066 Google Scholar
  7. Burkovics P, Hajdu I, Szukacsov V, Unk I, Haracska L (2009) Role of PCNA-dependent stimulation of 3′-phosphodiesterase and 3′–5′ exonuclease activities of human Ape2 in repair of oxidative DNA damage. Nucleic Acids Res 37(13):4247–4255. doi: 10.1093/nar/gkp357 PubMedCentralPubMedCrossRefGoogle Scholar
  8. Campagnoli C, Roberts IA, Kumar S, Bennett PR, Bellantuono I, Fisk NM (2001) Identification of mesenchymal stem/progenitor cells in human first-trimester fetal blood, liver, and bone marrow. Blood 98(8):2396–2402PubMedCrossRefGoogle Scholar
  9. Chen LB, Jiang XB, Yang L (2004) Differentiation of rat marrow mesenchymal stem cells into pancreatic islet beta-cells. World J Gastroenterol WJG 10(20):3016–3020Google Scholar
  10. De Bari C, Dell’Accio F, Tylzanowski P, Luyten FP (2001) Multipotent mesenchymal stem cells from adult human synovial membrane. Arthritis Rheum 44(8):1928–1942. doi: 10.1002/1529-0131(200108)44:8<1928:AID-ART331>3.0.CO;2-P PubMedCrossRefGoogle Scholar
  11. De Coppi P, Bartsch G Jr, Siddiqui MM, Xu T, Santos CC, Perin L, Mostoslavsky G, Serre AC, Snyder EY, Yoo JJ, Furth ME, Soker S, Atala A (2007) Isolation of amniotic stem cell lines with potential for therapy. Nat Biotechnol 25(1):100–106. doi: 10.1038/nbt1274 PubMedCrossRefGoogle Scholar
  12. Fink T, Abildtrup L, Fogd K, Abdallah BM, Kassem M, Ebbesen P, Zachar V (2004) Induction of adipocyte-like phenotype in human mesenchymal stem cells by hypoxia. Stem Cells 22(7):1346–1355. doi: 10.1634/stemcells.2004-0038 PubMedCrossRefGoogle Scholar
  13. Gerdes J, Lemke H, Baisch H, Wacker HH, Schwab U, Stein H (1984) Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67. J Immunol 133(4):1710–1715PubMedGoogle Scholar
  14. Gong X, Sun Z, Cui D, Xu X, Zhu H, Wang L, Qian W, Han X (2012) Isolation and characterization of lung resident mesenchymal stem cells capable of differentiating into alveolar epithelial type II cells. Cell Biol Int. doi: 10.1042/CBI20120073 Google Scholar
  15. Herrera MB, Bruno S, Buttiglieri S, Tetta C, Gatti S, Deregibus MC, Bussolati B, Camussi G (2006) Isolation and characterization of a stem cell population from adult human liver. Stem Cells 24(12):2840–2850. doi: 10.1634/stemcells.2006-0114 PubMedCrossRefGoogle Scholar
  16. Huss R, Lange C, Weissinger EM, Kolb HJ, Thalmeier K (2000) Evidence of peripheral blood-derived, plastic-adherent CD34(-/low) hematopoietic stem cell clones with mesenchymal stem cell characteristics. Stem Cells 18(4):252–260. doi: 10.1634/stemcells.18-4-252 PubMedCrossRefGoogle Scholar
  17. Jaiswal N, Haynesworth SE, Caplan AI, Bruder SP (1997) Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro. J Cell Biochem 64(2):295–312PubMedCrossRefGoogle Scholar
  18. Lange C, Bruns H, Kluth D, Zander AR, Fiegel HC (2006) Hepatocytic differentiation of mesenchymal stem cells in cocultures with fetal liver cells. World J Gastroenterol WJG 12(15):2394–2397Google Scholar
  19. Lee KD, Kuo TK, Whang-Peng J, Chung YF, Lin CT, Chou SH, Chen JR, Chen YP, Lee OK (2004) In vitro hepatic differentiation of human mesenchymal stem cells. Hepatology 40(6):1275–1284. doi: 10.1002/hep.20469 PubMedCrossRefGoogle Scholar
  20. Mackay AM, Beck SC, Murphy JM, Barry FP, Chichester CO, Pittenger MF (1998) Chondrogenic differentiation of cultured human mesenchymal stem cells from marrow. Tissue Eng 4(4):415–428PubMedCrossRefGoogle Scholar
  21. Maga G, Hubscher U (2003) Proliferating cell nuclear antigen (PCNA): a dancer with many partners. J Cell Sci 116(Pt 15):3051–3060. doi: 10.1242/jcs.00653 PubMedCrossRefGoogle Scholar
  22. Meirelles Lda S, Nardi NB (2003) Murine marrow-derived mesenchymal stem cell: isolation, in vitro expansion, and characterization. Br J Haematol 123(4):702–711PubMedCrossRefGoogle Scholar
  23. Miele A, Medina R, van Wijnen AJ, Stein GS, Stein JL (2007) The interactome of the histone gene regulatory factor HiNF-P suggests novel cell cycle related roles in transcriptional control and RNA processing. J Cell Biochem 102(1):136–148. doi: 10.1002/jcb.21284 PubMedCrossRefGoogle Scholar
  24. Moon JH, Lee JR, Jee BC, Suh CS, Kim SH, Lim HJ, Kim HK (2008) Successful vitrification of human amnion-derived mesenchymal stem cells. Hum Reprod 23(8):1760–1770. doi: 10.1093/humrep/den202 PubMedCrossRefGoogle Scholar
  25. Moreno R, Martinez-Gonzalez I, Rosal M, Farwati A, Gratacos E, Aran JM (2010) Characterization of mesenchymal stem cells isolated from the rabbit fetal liver. Stem cells Dev 19(10):1579–1588. doi: 10.1089/scd.2009.0514 PubMedCrossRefGoogle Scholar
  26. Piryaei A, Valojerdi MR, Shahsavani M, Baharvand H (2011) Differentiation of bone marrow-derived mesenchymal stem cells into hepatocyte-like cells on nanofibers and their transplantation into a carbon tetrachloride-induced liver fibrosis model. Stem cell Rev 7(1):103–118. doi: 10.1007/s12015-010-9126-5 PubMedCrossRefGoogle Scholar
  27. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284(5411):143–147PubMedCrossRefGoogle Scholar
  28. Sanchez-Ramos J, Song S, Cardozo-Pelaez F, Hazzi C, Stedeford T, Willing A, Freeman TB, Saporta S, Janssen W, Patel N, Cooper DR, Sanberg PR (2000) Adult bone marrow stromal cells differentiate into neural cells in vitro. Exp Neurol 164(2):247–256. doi: 10.1006/exnr.2000.7389 PubMedCrossRefGoogle Scholar
  29. Strem BM, Hicok KC, Zhu M, Wulur I, Alfonso Z, Schreiber RE, Fraser JK, Hedrick MH (2005) Multipotential differentiation of adipose tissue-derived stem cells. Keio J Med 54(3):132–141PubMedCrossRefGoogle Scholar
  30. Tamir A, Petrocelli T, Stetler K, Chu W, Howard J, Croix BS, Slingerland J, Ben-David Y (2000) Stem cell factor inhibits erythroid differentiation by modulating the activity of G1-cyclin-dependent kinase complexes: a role for p27 in erythroid differentiation coupled G1 arrest. Cell Growth Differ Mol Biol J Am Assoc Cancer Res 11(5):269–277Google Scholar
  31. Travali S, Ku DH, Rizzo MG, Ottavio L, Baserga R, Calabretta B (1989) Structure of the human gene for the proliferating cell nuclear antigen. J Biol Chem 264(13):7466–7472PubMedGoogle Scholar
  32. Wakitani S, Saito T, Caplan AI (1995) Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-azacytidine. Muscle Nerve 18(12):1417–1426. doi: 10.1002/mus.880181212 PubMedCrossRefGoogle Scholar
  33. Worster AA, Brower-Toland BD, Fortier LA, Bent SJ, Williams J, Nixon AJ (2001) Chondrocytic differentiation of mesenchymal stem cells sequentially exposed to transforming growth factor-beta1 in monolayer and insulin-like growth factor-I in a three-dimensional matrix. J Orthopaedic Res 19(4):738–749. doi: 10.1016/S0736-0266(00)00054-1 CrossRefGoogle Scholar
  34. Yoshimura H, Muneta T, Nimura A, Yokoyama A, Koga H, Sekiya I (2007) Comparison of rat mesenchymal stem cells derived from bone marrow, synovium, periosteum, adipose tissue, and muscle. Cell Tissue Res 327(3):449–462. doi: 10.1007/s00441-006-0308-z PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Taofeng Lu
    • 1
  • Pengfei Hu
    • 1
  • Xiaohua Su
    • 1
  • Changli Li
    • 1
  • Yuehui Ma
    • 1
  • Weijun Guan
    • 1
  1. 1.Institute of Animal SciencesChinese Academy of Agricultural SciencesBeijingChina

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