World Journal of Urology

, 26:341 | Cite as

Bone marrow stem cells for urologic tissue engineering

  • Dave Shukla
  • Geoffrey N. Box
  • Robert A. Edwards
  • Darren R. Tyson
Topic Paper



Experiments in rats and dogs have demonstrated the potential of bone marrow-derived mesenchymal stem cells (MSCs) for urinary tract tissue engineering. However, the small graft size in rats and a failure to identify the MSCs in engineered tissues made it difficult to assess the true potential of these cells. Our goals were to characterize MSCs from pigs, determine their ability to differentiate into smooth muscle cells (SMCs) and use them in an autologous augmentation cystoplasty.


MSCs were isolated from pigs and analyzed for common markers of MSCs by flow cytometry. SMC differentiation was determined by immunoblotting. MSCs were isolated, genetically labeled, expanded in vitro, seeded onto small intestinal submucosa (SIS) and used for autologous bladder augmentation.


Porcine MSCs are morphologically and immunophenotypically similar to human MSCs. Culturing MSCs at low density enhances proliferation rates. MSCs consistently differentiate into mature SMCs in vitro when maintained at confluence. Labeled MSCs grew on SIS over one week in vitro and survived a 2-week implantation as an autologous bladder augment in vivo. Some label-positive cells with SMC morphology were detected, but most SMCs were negative. Notably, many cells with a urothelial morphology stained positively.


Porcine MSCs have similar properties to MSCs from other species and consistently undergo differentiation into mature SMC in vitro under specific culture conditions. Labeled MSCs within SIS may assist tissue regeneration in augmentation cystoplasty but may not significantly incorporate into smooth muscle bundles.


Bone marrow Mesenchymal stem cells Multipotent stromal cells Tissue engineering Regenerative medicine 



We gratefully acknowledge Umesh Patel and Cook Biotech for providing SurgiSIS. We would also like to thank Ralph Clayman for his support, Lorena Andrade and Reza Alipanah for their assistance with the animal studies, and Alice Lau for her assistance with the immunoblotting and proliferation assays.

Conflict of interest statement

There is no conflict of interest.


  1. 1.
    Atala A (2000) Tissue engineering for bladder substitution. World J Urol 18:364–370PubMedCrossRefGoogle Scholar
  2. 2.
    Zhang Y, Kropp BP, Moore P, Cowan R, Furness PD 3rd, Kolligian ME et al (2000) Coculture of bladder urothelial and smooth muscle cells on small intestinal submucosa: potential applications for tissue engineering technology. J Urol 164:928–934; discussion 934–5PubMedCrossRefGoogle Scholar
  3. 3.
    Kropp BP, Cheng EY (2000) Bioengineering organs using small intestinal submucosa scaffolds: in vivo tissue-engineering technology. J Endourol 14:59–62PubMedCrossRefGoogle Scholar
  4. 4.
    Sievert KD, Tanagho EA (2000) Organ-specific acellular matrix for reconstruction of the urinary tract. World J Urol 18:19–25PubMedCrossRefGoogle Scholar
  5. 5.
    Hiraga S, Iida T, Kitamura M, Takamiya T, Wakabayashi T, Hida M et al (1989) Experimental study of urinary vesical transplantation. Transplant Proc 21:3194–3196PubMedGoogle Scholar
  6. 6.
    Takeuchi K, Ohoka H, Yokoyama M, Iwata H, Takeuchi M (1996) A technique of urinary bladder transplantation in the rat. Transplant Proc 28:1978–1979PubMedGoogle Scholar
  7. 7.
    Takeuchi K, Takechi S, Ohoka H, Yokoyama M, Iwata H, Takeuchi M et al (1997) Histological study of urinary bladder transplantation in rats. Transplantation 63:922–926PubMedCrossRefGoogle Scholar
  8. 8.
    Baksh D, Song L, Tuan RS (2004) Adult mesenchymal stem cells: characterization, differentiation, and application in cell and gene therapy. J Cell Mol Med 8:301–316PubMedCrossRefGoogle Scholar
  9. 9.
    Eckfeldt CE, Mendenhall EM, Verfaillie CM (2005) The molecular repertoire of the ‘Almighty’ stem cell. Nat Rev Mol Cell Biol 6:726–737PubMedCrossRefGoogle Scholar
  10. 10.
    Prockop DJ, Gregory CA, Spees JL (2003) One strategy for cell and gene therapy: harnessing the power of adult stem cells to repair tissues. Proc Natl Acad Sci USA 100(Suppl 1):11917–11923PubMedCrossRefGoogle Scholar
  11. 11.
    Charbord P, Lerat H, Newton I, Tamayo E, Gown AM, Singer JW et al (1990) The cytoskeleton of stromal cells from human bone marrow cultures resembles that of cultured smooth muscle cells. Exp Hematol 18:276–282PubMedGoogle Scholar
  12. 12.
    Charbord P, Tamayo E, Deschaseaux F, Remy-Martin JP, Pelletier L, Sensebe L et al (1999) The hematopoietic microenvironment: phenotypic and functional characterization of human marrow vascular stromal cells. Hematology 4:257–282PubMedGoogle Scholar
  13. 13.
    Galmiche MC, Koteliansky VE, Briere J, Herve P, Charbord P (1993) Stromal cells from human long-term marrow cultures are mesenchymal cells that differentiate following a vascular smooth muscle differentiation pathway. Blood 82:66–76PubMedGoogle Scholar
  14. 14.
    Li J, Sensebe L, Herve P, Charbord P (1995) Nontransformed colony-derived stromal cell lines from normal human marrows. II. Phenotypic characterization and differentiation pathway. Exp Hematol 23:133–141PubMedGoogle Scholar
  15. 15.
    Chung SY, Krivorov NP, Rausei V, Thomas L, Frantzen M, Landsittel D et al (2005) Bladder reconstitution with bone marrow derived stem cells seeded on small intestinal submucosa improves morphological and molecular composition. J Urol 174:353–359PubMedCrossRefGoogle Scholar
  16. 16.
    Zhang Y, Lin HK, Frimberger D, Epstein RB, Kropp BP (2005) Growth of bone marrow stromal cells on small intestinal submucosa: an alternative cell source for tissue engineered bladder. BJU Int 96:1120–1125PubMedCrossRefGoogle Scholar
  17. 17.
    Vacanti V, Kong E, Suzuki G, Sato K, Canty JM, Lee T (2005) Phenotypic changes of adult porcine mesenchymal stem cells induced by prolonged passaging in culture. J Cell Physiol 205:194–201PubMedCrossRefGoogle Scholar
  18. 18.
    Zhang Y, Frimberger D, Cheng EY, Lin HK, Kropp BP (2006) Challenges in a larger bladder replacement with cell-seeded and unseeded small intestinal submucosa grafts in a subtotal cystectomy model. BJU Int 98:1100–1105PubMedCrossRefGoogle Scholar
  19. 19.
    Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD et al (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147PubMedCrossRefGoogle Scholar
  20. 20.
    Suva D, Garavaglia G, Menetrey J, Chapuis B, Hoffmeyer P, Bernheim L et al (2004) Non-hematopoietic human bone marrow contains long-lasting, pluripotential mesenchymal stem cells. J Cell Physiol 198:110–118PubMedCrossRefGoogle Scholar
  21. 21.
    Lodie TA, Blickarz CE, Devarakonda TJ, He C, Dash AB, Clarke J et al (2002) Systematic analysis of reportedly distinct populations of multipotent bone marrow-derived stem cells reveals a lack of distinction. Tissue Eng 8:739–751PubMedCrossRefGoogle Scholar
  22. 22.
    Vázquez ME, Cabarcos MR, Román TD, Stein AJ, García ND, Nazar BA et al (2005) Cellular cardiomyoplasty: development of a technique to culture human myoblasts for clinical transplantation. Cell Tissue Bank 6:117–124PubMedCrossRefGoogle Scholar
  23. 23.
    Colter DC, Sekiya I, Prockop DJ (2001) Identification of a subpopulation of rapidly self-renewing and multipotential adult stem cells in colonies of human marrow stromal cells. Proc Natl Acad Sci USA 98:7841–7845PubMedCrossRefGoogle Scholar
  24. 24.
    Owens GK, Kumar MS, Wamhoff BR (2004) Molecular regulation of vascular smooth muscle cell differentiation in development and disease. Physiol Rev 84:767–801PubMedCrossRefGoogle Scholar
  25. 25.
    Kanematsu A, Yamamoto S, Iwai-Kanai E, Kanatani I, Imamura M, Adam RM et al (2005) Induction of smooth muscle cell-like phenotype in marrow-derived cells among regenerating urinary bladder smooth muscle cells. Am J Pathol 166:565–573PubMedGoogle Scholar
  26. 26.
    Kashiwada K, Nishida W, Hayashi K, Ozawa K, Yamanaka Y, Saga H et al (1997) Coordinate expression of alpha-tropomyosin and caldesmon isoforms in association with phenotypic modulation of smooth muscle cells. J Biol Chem 272:15396–15404PubMedCrossRefGoogle Scholar
  27. 27.
    Ross JJ, Hong Z, Willenbring B, Zeng L, Isenberg B, Lee EH et al (2006) Cytokine-induced differentiation of multipotent adult progenitor cells into functional smooth muscle cells. J Clin Invest 116:3139–3149PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Dave Shukla
    • 1
  • Geoffrey N. Box
    • 2
  • Robert A. Edwards
    • 3
  • Darren R. Tyson
    • 1
  1. 1.Department of UrologyUniversity of California IrvineOrangeUSA
  2. 2.Department of UrologyUniversity of California IrvineOrangeUSA
  3. 3.Department of PathologyUniversity of California IrvineIrvineUSA

Personalised recommendations