Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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In Vitro Differentiation of Mesenchymal Progenitor Cells Derived from Porcine Umbilical Cord Blood

  • Kumar, Basavarajappa Mohana (College of Veterinary Medicine, Gyeongsang National University) ;
  • Yoo, Jae-Gyu (College of Veterinary Medicine, Gyeongsang National University) ;
  • Ock, Sun-A (College of Veterinary Medicine, Gyeongsang National University) ;
  • Kim, Jung-Gon (College of Veterinary Medicine, Gyeongsang National University) ;
  • Song, Hye-Jin (College of Veterinary Medicine, Gyeongsang National University) ;
  • Kang, Eun-Ju (College of Veterinary Medicine, Gyeongsang National University) ;
  • Cho, Seong-Keun (Division of Applied Life Science, Gyeongsang National University) ;
  • Lee, Sung-Lim (College of Veterinary Medicine, Gyeongsang National University) ;
  • Cho, Jae-Hyeon (College of Veterinary Medicine, Gyeongsang National University) ;
  • Balasubramanian, Sivasankaran (College of Veterinary Medicine, Gyeongsang National University) ;
  • Rho, Gyu-Jin (College of Veterinary Medicine, Gyeongsang National University)
  • Received : 2007.04.23
  • Accepted : 2007.07.03
  • Published : 2007.12.31
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Abstract

Mesenchymal stem/progenitor cells (MPCs) were isolated from porcine umbilical cord blood (UCB) and their morphology, proliferation, cell cycle status, cell-surface antigen profile and expression of hematopoietic cytokines were characterized. Their capacity to differentiate in vitro into osteocytes, adipocytes and chondrocytes was also evaluated. Primary cultures of adherent porcine MPCs (pMPCs) exhibited a typical fibroblast-like morphology with significant renewal capacity and proliferative ability. Subsequent robust cell growth was indicated by the high percentage of quiescent (G0/G1) cells. The cells expressed the mesenchymal surface markers, CD29, CD49b and CD105, but not the hematopoietic markers, CD45 and CD133 and synthesized hematopoietic cytokines. Over 21 days of induction, the cells differentiated into osteocytes adipocytes and chondrocytes. The expression of lineage specific genes was gradually upregulated during osteogenesis, adipogenesis and chondrogenesis. We conclude that porcine umbilical cord blood contains a population of MPCs capable of self-renewal and of differentiating in vitro into three classical mesenchymal lineages.

Keywords

References

  1. Bosch, P., Pratt, S. L., and Stice, S. L. (2006) Isolation, characterization, gene modification, and nuclear reprogramming of porcine mesenchymal stem cells. Biol. Reprod. 74, 46-57 https://doi.org/10.1095/biolreprod.105.045138
  2. Bosnakovski, D., Mizuno, M., Kim, G., Takagi, S., Okumura, M., et al. (2005) Isolation and multilineage differentiation of bovine bone marrow mesenchymal stem cells. Cell Tissue Res. 319, 243-253 https://doi.org/10.1007/s00441-004-1012-5
  3. Carlin, R., Davis, D., Weiss, M., Schultz, B., and Troyer, D. (2006) Expression of early transcription factors Oct-4, Sox-2 and Nanog by porcine umbilical cord (PUC) matrix cells. Reprod. Biol. Endocinol. 4, 8 doi: 10.1186/1477-7827-4-8
  4. Chang, Y. J., Tseng, C. P., Hsu, L. F., Hsieh, T. B., and Hwang, S. M. (2006) Characterization of two populations of mesenchymal progenitor cells in umbilical cord blood. Cell Biol. Intl. 30, 495-499 https://doi.org/10.1016/j.cellbi.2005.12.009
  5. Colleoni, S., Donofrio, G., Lagutina, I., Duchi, R., Galli, C., et al. (2005) Establishment, differentiation, electroporation, viral transduction, and nuclear transfer of bovine and porcine mesenchymal stem cells. Cloning Stem Cells 7, 154-166 https://doi.org/10.1089/clo.2005.7.154
  6. De Boer, J., Wang, H. J., and Van Blitterswijk, C. (2004) Effects of Wnt signaling on proliferation and differentiation of human mesenchymal stem cells. Tissue Eng. 10, 393-401 https://doi.org/10.1089/107632704323061753
  7. Erices, A., Conget, P., and Minguell, J. J. (2000) Mesenchymal progenitor cells in human umbilical cord blood. Br. J. Haematol. 109, 235-242 https://doi.org/10.1046/j.1365-2141.2000.01986.x
  8. Faast, R., Harrison, S. J., Beebe, L. F., Mcilfatrick, S. M., Ashman, R. J., et al. (2006) Use of adult mesenchymal stem cells isolated from bone marrow and blood for somatic cell nuclear transfer in pigs. Cloning Stem Cells 8, 166-173 https://doi.org/10.1089/clo.2006.8.166
  9. Fuchs, J. R., Hannouche, D., Terada, S., Zand, S., Vacanti, J. P., et al. (2005) Cartilage engineering from ovine umbilical cord blood mesenchymal progenitor cells. Stem Cells 23, 958-964 https://doi.org/10.1634/stemcells.2004-0310
  10. Goodwin, H. S., Bicknese, A. R., Chien, S. N., Bogucki, B. D., Quinn, C. O., et al. (2001) Multilineage differentiation activity by cells isolated from umbilical cord blood expression of bone, fat, and neural markers. Biol. Blood Marrow Transplant. 7, 581-588 https://doi.org/10.1053/bbmt.2001.v7.pm11760145
  11. Gregoire, F. M., Smas, C. M., and Sul, H. S. (1998) Understanding adipocyte differentiation. Physiol. Rev. 78, 783-809 https://doi.org/10.1152/physrev.1998.78.3.783
  12. Jager, M., Bachmann, R., Scharfstadt, A., and Krauspe, R. (2006) Ovine cord blood accommodates multipotent mesenchymal progenitor cells. In Vivo 20, 205-214
  13. Jeong, J. A., Hong, S. H., Gang, E. J., Ahn, C., Hwang, S. H., et al. (2005) Differential gene expression profiling of human umbilical cord blood-derived mesenchymal stem cells. Stem Cells 23, 584-593 https://doi.org/10.1634/stemcells.2004-0304
  14. Jin, H. F., Mohana kumar, B., Kim, J. G., Song, H. J., Jeong, Y. J., et al. (2007) Enhanced development of porcine embryos cloned from bone marrow mesenchymal stem cells. Intl. J. Dev. Biol. 50, 85-90
  15. Johnstone, B., Hering, T. M., Caplan, A. I., Goldberg, V. M., and Yoo, J. U. (1998) In vitro chondrogenesis of bone marrow–. derived mesenchymal progenitor cells. Exp. Cell Res. 238, 265-272 https://doi.org/10.1006/excr.1997.3858
  16. Kern, S., Eichler, H., Stoeve, J., Kluter, H., and Bieback, K. (2006) Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue stem cells. Stem Cells 24, 1294-1301 https://doi.org/10.1634/stemcells.2005-0342
  17. Lee, O. K., Kuo, T. K., Chen, W. M., Lee, K. D., Hsieh, S. L., et al. (2004) Isolation of multipotent mesenchymal stem cells from umbilical cord blood. Blood 103, 1669-1675 https://doi.org/10.1182/blood-2003-05-1670
  18. Lu, L. L., Liu, Y. J., Yang, S. G., Zhao, Q. J., Wang, X., et al. (2006) Isolation and characterization of human umbilical cord mesenchymal stem cells with hematopoiesis-supportive function and other potentials. Haematologica 91, 1017-1026
  19. Mastrogiacomo, M., Cancedda, R., and Quarto, R. (2001) Effect of different growth factors on the chondrogenic potential of human bone marrow stromal cells. Osteoarthritis Cartilage 9, S36-S40
  20. McBeath, R., Pirone, D. M., Nelson, C. M., Bhadriraju, K., and Chen, C. S. (2004) Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev. Cell 6, 483-495 https://doi.org/10.1016/S1534-5807(04)00075-9
  21. Minguell, J. J., Erices, A., and Conget, P. (2001) Mesenchymal stem cells. Exp. Biol. Med. 226, 507-520 https://doi.org/10.1177/153537020122600603
  22. Naruse, K., Urabe, K., Mukaida, T., Ueno, T., Migishima, F., et al. (2004) Spontaneous differentiation of mesenchymal stem cells obtained from fetal rat circulation. Bone 35, 850-858 https://doi.org/10.1016/j.bone.2004.05.006
  23. Pittenger, M. F., Mackay, A. M., Beck, S. C., Jaiswal, R. K., Douglas, R., et al. (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284, 143-147 https://doi.org/10.1126/science.284.5411.143
  24. Price, E. M., Prather, R. S., and Foley, C. M. (2006) Multipotent adult progenitor cell lines originating from the peripheral blood of green fluorescent protein transgenic swine. Stem Cells Dev. 15, 507-522 https://doi.org/10.1089/scd.2006.15.507
  25. Ringe, J., Kaps, C., Schmitt, B., Buscher, K., Bartel, J., et al. (2002) Porcine mesenchymal stem cells: induction of distinct mesenchymal cell lineages. Cell Tissue Res. 307, 321-327 https://doi.org/10.1007/s00441-002-0525-z
  26. Rogers, I. and Casper, R. F. (2004) Umbilical cord blood stem cells. Best Pract. Res. Clin. Obstet. Gynaecol. 18, 893-908 https://doi.org/10.1016/j.bpobgyn.2004.06.004
  27. Sergeeva, N. S., Sviridova, I. K., Kirsanova, V. A., Akhmedova, S. A., Marshutina, N. V., et al. (2006) Study of growth parameters and potentialities of differentiation of multipotent mesenchymal stromal cells from rat bone marrow in vitro. Bull. Exp. Biol. Med. 141, 530-535 https://doi.org/10.1007/s10517-006-0214-0
  28. Tropel, P., Noel, D., Platet, N., Legrand, F., Benabid, A., et al. (2004) Isolation and characterization of mesenchymal stem cells from adult mouse bone marrow. Exp. Cell Res. 295, 395-406 https://doi.org/10.1016/j.yexcr.2003.12.030
  29. Vacanti, V., Kong, E., Suzuki, G., Sato, K., Canty, J. M., et al. (2005) Phenotypic changes of adult porcine mesenchymal stem cells induced by prolonged passaging in culture. J. Cell. Physiol. 205, 194-201 https://doi.org/10.1002/jcp.20376
  30. Vidal, M. A., Kilroy, G. E., Johnson, J. R., Lopes, M. J., Moore, R. M., et al. (2006) Cell growth characteristics and differentiation frequency of adherent equine bone marrow-derived mesenchymal stromal cells: adipogenic and osteogenic capacity. Vet. Surg. 35, 601-610 https://doi.org/10.1111/j.1532-950X.2006.00197.x
  31. Wagner, W., Wein, F., Seckinger, A., Frankhauser, M., Wirkner, U., et al. (2005) Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood. Exp. Hematol. 33, 1402-1416 https://doi.org/10.1016/j.exphem.2005.07.003
  32. Worster, A. A., Nixon, A. J., Brower-Toland, B. D., and Williams, J. (2000) Effect of transforming growth factor beta 1 on chondrogenic differentiation of cultured equine mesenchymal stem cells. Am. J. Res. 61, 1003-1010 https://doi.org/10.2460/ajvr.2000.61.1003
  33. Zeng, L., Rahrmann, E., Hu, Q., Lund, T., Sandquist, L., et al. (2006) Multipotent adult progenitor cells from swine bone marrow. Stem Cells 24, 2355-2366 https://doi.org/10.1634/stemcells.2005-0551