Human adipose-derived stem cells for the treatment of intracerebral hemorrhage in rats via femoral intravenous injection

Abstract

Human adipose-derived stem cells (huADSC) were generated from fat tissue of a 65-year-old male donor. Flow cytometry and reverse transcription polymerase chain reaction (RT-PCR) analyses indicated that the huADSC express neural cell proteins (MAP2, GFAP, nestin and β-III tubulin), neurotrophic growth factors (BDNF and GDNF), and the chemotactic factor CXCR4 and its corresponding ligand CXCL12. In addition, huADSC expressed the characteristic mesenchymal stem cell (MSC) markers CD29, CD44, CD73, CD90, CD105 and HLA class I. The huADSC were employed, via a right femoral vein injection, to treat rats inflicted with experimental intracerebral hemorrhage (ICH). Behavioral measurement on the experimental animals, seven days after the huADSC therapy, showed a significant functional improvement in the rats with stem cell therapy in comparison with rats of the control group without the stem cell therapy. The injected huADSC were detectable in the brains of the huADSC treated rats as determined by histochemistry analysis, suggesting a role of the infused huADSC in facilitating functional recovery of the experimental animals with ICH induced stroke.

Abbreviations

ADAS:

adipose-derived adult stem

ADSC:

adipose-derived stem cell

FCS:

fetal calf serum

hAdMSC:

adipose derived mesenchymal stem cell

hASC:

adipose tissue-derived stem cell

hATSC:

adipose tissue stromal cell

ICH:

intracerebral hemorrhage

MCAO:

middle cerebral artery occlusion

MSC:

mesenchymal stem cell

SVF:

stromal-vascular fraction

References

  1. 1.

    Xi, G., Keep, R.F. and Hoff, J.T. Mechanisms of brain injury after intracerebral haemorrhage. Lancet Neurol. 53 (2006) 53–63.

    Article  Google Scholar 

  2. 2.

    Broderick, J.P., Adams, H.P. Jr., Barsan, W., Feinberg, W., Feldmann, E., Grotta, J., Kase, C., Krieger, D., Mayberg, M., Tilley, B., Zabramski, J.M. and Zuccarello, M. Guidelines for the management of spontaneous intracerebral hemorrhage: A statement for healthcare professionals from a special writing group of the Stroke Council, American Heart Association. Stroke, 30 (1999) 905–915.

    PubMed  Article  CAS  Google Scholar 

  3. 3.

    Mendelow, A.D., Gregson, B.A., Fernandes, H.M., Murray, G.D., Teasdale, G.M., Hope, D.T., Karimi, A., Shaw, M.D., Barer, D.H. and STICH investigators. Early surgery versus initial conservative treatment in patients with spontaneous supratentorial intracerebral haematomas in the International Surgical Trial in Intracerebral Haemorrhage (STICH): a randomised trial. Lancet, 365 (2005) 387–397.

    PubMed  Google Scholar 

  4. 4.

    Del Bigio, M.R., Yan, H.J., Buist, R. and Peeling, J. Experimental intracerebral hemorrhage in rats. Magnetic resonance imaging and histopathological correlates. Stroke, 27 (1996) 2312–2319; discussion 2319–2320.

    PubMed  Article  Google Scholar 

  5. 5.

    Gong, C., Hoff, J.T. and Keep, R.F. Acute inflammatory reaction following experimental intracerebral hemorrhage in rat. Brain Res. 871 (2000) 57–65.

    PubMed  Article  CAS  Google Scholar 

  6. 6.

    Xue, M. and Del Bigio, M.R. Intracerebral injection of autologous whole blood in rats: time course of inflammation and cell death. Neurosci. Lett. 283 (2000) 230–232.

    PubMed  Article  CAS  Google Scholar 

  7. 7.

    Fenstermacher, J.D., Knight, R.A., Ewing, J.R., Nagaraja, T., Nagesh, V., Yee, J.S. and Arniego, P.A. Estimating blood-brain barrier opening in a rat model of hemorrhagic transformation with Patlak plots of Gd-DTPA contrast-enhanced MRI. Acta Neurochir. Suppl. 86 (2003) 35–37.

    PubMed  Article  CAS  Google Scholar 

  8. 8.

    Hoff, J.T. and Xi, G. Brain edema from intracerebral hemorrhage. Acta Neurochir. Suppl. 86 (2003) 11–15.

    PubMed  Article  CAS  Google Scholar 

  9. 9.

    Kitaoka, T., Hua, Y., Xi, G., Hoff, J.T. and Keep, R.F. Delayed argatroban treatment reduces edema in a rat model of intracerebral hemorrhage. Stroke 33 (2002) 3012–3018.

    PubMed  Article  CAS  Google Scholar 

  10. 10.

    MacLellan, C.L., Silasi, G., Poon, C.C., Edmundson, C.L., Buist, R., Peeling, J. and Colbourne, F. Intracerebral hemorrhage models in rat: comparing collagenase to blood infusion. J. Cereb. Blood Flow Metab. 28 (2008) 516–525.

    PubMed  Article  CAS  Google Scholar 

  11. 11.

    Rincon, F. and Mayer, S.A. Novel therapies for intracerebral hemorrhage. Curr. Opin. Crit. Care, 10 (2004) 94–100.

    PubMed  Article  Google Scholar 

  12. 12.

    Wu, J., Hua, Y., Keep, R.F., Nakamura, T., Hoff, J.T. and Xi, G. Iron and iron-handling proteins in the brain after intracerebral hemorrhage. Stroke, 34 (2003) 2964–2969.

    PubMed  Article  CAS  Google Scholar 

  13. 13.

    Bai, X., Ma, J., Pan, Z., Song, Y.H., Freyberg, S., Yan, Y., Vykoukal, D. and Alt, E. Electrophysiological properties of human adipose tissue-derived stem cells. Am. J. Physiol. Cell Physiol. 293 (2007) C1539–C1550.

    PubMed  Article  CAS  Google Scholar 

  14. 14.

    Bunnell, B.A., Flaat, M., Gagliardi, C., Patel, B. and Ripoll, C. Adiposederived stem cells: isolation, expansion and differentiation. Methods, 45 (2008) 115–120.

    PubMed  Article  CAS  Google Scholar 

  15. 15.

    Gimble, J.M., Katz, A.J. and Bunnell, B.A. Adipose-derived stem cells for regenerative medicine. Circ. Res. 100 (2007) 1249–1260.

    PubMed  Article  CAS  Google Scholar 

  16. 16.

    Hong, L., Peptan, I.A., Colpan, A. and Daw, J.L. Adipose tissue engineering by human adipose-derived stromal cells. Cells Tissues Organs, 183 (2006) 133–140.

    PubMed  Article  CAS  Google Scholar 

  17. 17.

    Kang, S.K., Lee, D.H., Bae, Y.C., Kim, H.K., Baik, S.Y. and Jung, J.S. Improvement of neurological deficits by intracerebral transplantation of human adipose tissue-derivrd stromal cells after cerebral ischemia in rats. Exp. Neurol. 183 (2003) 355–366.

    PubMed  Article  CAS  Google Scholar 

  18. 18.

    Lin, S.D., Wang, K.H. and Kao, A.P. Engineered adipose tissue of predefined shape and dimensions from human adipose-derived mesenchymal stem cells. Tissue Eng. Part A, 14 (2008) 571–581.

    PubMed  Article  CAS  Google Scholar 

  19. 19.

    Witkowska-Zimny, M. and Walenko, K. Stem cells from adipose tissue. Cell. Mol. Biol. Lett. 16 (2011) 236–257.

    PubMed  Article  Google Scholar 

  20. 20.

    Ohta, Y., Takenaga, M., Tokura, Y., Hamaguchi, A., Matsumoto, T., Kano, K., Mugishima, H., Okano, H. and Igarashi, R. Mature adipocyte-derived cells, dedifferentiated fat cells (DFAT), promoted functional recovery from spinal cord injury-induced motor dysfunction in rats. Cell Transplant. 17 (2008) 877–886.

    PubMed  Article  Google Scholar 

  21. 21.

    Safford, K.M., Hicok, K.C., Safford, S.D., Halvorsen, Y.D., Wilkison, W.O., Gimble, J.M. and Rice, H.E. Neurogenic differentiation of murine and human adipose-derived stromal cells. Biochem. Biophy. Res. Commun. 294 (2002) 371–379.

    Article  CAS  Google Scholar 

  22. 22.

    Planat-Bénard, V., Menard, C., André, M., Puceat, M., Perez, A., Garcia-Verdugo, J.M., Pénicaud, L., Casteilla, L. Spontaneous cardiomyocyte differentiation from adipose tissue stroma cells. Circ. Res. 94 (2004) 223–229.

    PubMed  Article  Google Scholar 

  23. 23.

    Kim, J.M., Lee, S.T., Chu, K., Jung, K.H., Song, E.C., Kim, S.J., Sinn, D.I., Kim, J.H., Park, D.K., Kang, K.M., Hyung Hong, N., Park, H.K., Won, C.H., Kim, K.H., Kim, M., Kun Lee, S. and Roh, J.K. Systemic transplantation of human adipose stem cells attenuated cerebral inflammation and degeneration in a hemorrhagic stroke model. Brain Res. 1183 (2007) 43–50.

    PubMed  Article  CAS  Google Scholar 

  24. 24.

    Yang, K.L., Chen, M.F., Liao, C.H., Pang, C.Y. and Lin, P.Y.. A simple and efficient method for generating Nurr1-positive neuronal stem cells from human wisdom teeth (tNSC) and the potential of tNSC for stroke therapy. Cytotherapy, 11 (2009) 606–617.

    PubMed  Article  CAS  Google Scholar 

  25. 25.

    Rosenberg, G.A., Mun-Bryce, S., Wesley, M. and Kornfeld, M. Collagenase-induced intracerebral hemorrhage in rats. Stroke, 21 (1990) 801–807.

    PubMed  Article  CAS  Google Scholar 

  26. 26.

    MacLellan, C.L., Silasi, G., Poon, C.C., Edmundson, C.L., Buist, R., Peeling, J. and Colbourne, F. Intracerebral hemorrhage models in rat: comparing collagenase to blood infusion. J. Cereb. Blood Flow Metab. 28 (2008) 516–525.

    PubMed  Article  CAS  Google Scholar 

  27. 27.

    Chen, J., Li, Y., Wang, L., Zhang, Z., Lu, D., Lu, M. and Chopp, M. Therapeutic benefit of intravenous administration of bone marrow stromal cells after cerebral ischemia in rats. Stroke, 32 (2001) 1005–1011.

    PubMed  Article  CAS  Google Scholar 

  28. 28.

    Witkowska-Zimny, M. and Wrobel, E. Perinatal sources of mesenchymal stem cells: Wharton’s jelly, amnion and chorion. Cell. Mol. Biol. Lett. 16 (2011) 493–514.

    PubMed  Article  Google Scholar 

  29. 29.

    Klassen, H., Schwartz, M.R., Bailey, A.H. and Young, M.J. Surface markers expressed by multipotent human and mouse neural progenitor cells include tetraspanins and non-protein epitopes. Neurosci. Lett. 312 (2001) 180–182.

    PubMed  Article  CAS  Google Scholar 

  30. 30.

    Zuk, P.A., Zhu, M., Ashjian, P., De Ugarte, D.A., Huang, J.I., Mizuno, H., Alfonso, Z.C., Fraser, J.K., Benhaim, P. and Hedrick, M.H. Human adipose tissue is a source of multipotent stem cells. Mol. Biol. Cell. 13 (2002) 4279–4295.

    PubMed  Article  CAS  Google Scholar 

  31. 31.

    Chen, L., He, D.M. and Zhang, Y. The differentiation of human placentaderived mesenchymal stem cells into dopaminergic cells in vitro. Cell. Mol. Biol. Lett. 14 (2009) 528–536.

    PubMed  Article  CAS  Google Scholar 

  32. 32.

    Mahmood, A., Lu, D., Wang, L. and Chopp, M. Intracerebral transplantation of marrow stromal cells cultured with neurotrophic factors promotes functional recovery in adult rats subjected to traumatic brain injury. J. Neurotrauma, 19 (2002) 1609–1617.

    PubMed  Article  Google Scholar 

  33. 33.

    Nakagami, H., Maeda, K., Morishita, R., Iguchi, S., Nishikawa, T., Takami, Y., Kikuchi, Y., Saito, Y., Tamai, K., Ogihara, T. and Kaneda, Y. Novel autologous cell therapy in ischemic limb disease through growth factor secretion by cultured adipose tissue-derived stromal cells. Arterioscler. Thromb. Vasc. Biol. 25 (2005) 2542–2547.

    PubMed  Article  CAS  Google Scholar 

  34. 34.

    Tate, C.C., Fonck, C., McGrogan, M. and Case, C.C. Human mesenchymal stromal cells and their derivative, SB623 cells, rescue neural cells via trophic support following in vitro ischemia. Cell Transplant. 19 (2010) 973–984.

    PubMed  Article  Google Scholar 

  35. 35.

    Harting, M.T., Jimenez, F., Xue, H., Fischer, U.M., Baumgartner, J., Dash, P.K. and Cox, C.S. Intravenous mesenchymal stem cell therapy for traumatic brain injury. J. Neurosurg. 110 (2009) 1189–1197.

    PubMed  Article  CAS  Google Scholar 

  36. 36.

    Schrepfer, S., Deuse, T., Reichenspurner, H., Fischbein, M.P., Robbins, R.C., Pelletier, M.P. Stem cell transplantation: the lung barrier. Transplant. Proc. 39 (2007) 573–576.

    PubMed  Article  CAS  Google Scholar 

  37. 37.

    Mahmood, A., Lu, D. and Chopp, M. Intravenous administration of marrow stromal cells (MSCs) increases the expression of growth factors in rat brain after traumatic brain injury. J. Neurotrauma, 21 (2004) 33–39.

    PubMed  Article  Google Scholar 

  38. 38.

    Lin, G., Yang, R., Banie, L., Wang, G., Ning, H., Li, L.C., Lue, T.F., Lin, C.S. Effects of transplantation of adipose tissue-derived stem cells on prostate tumor. Prostate, 70 (2010) 1066–1073.

    PubMed  Article  Google Scholar 

  39. 39.

    Chung, S., Sonntag, K.C., Andersson, T., Bjorklund, L.M., Park, J.J., Kim, D.W., Kang, U.J., Isacson, O. and Kim, K.S. Genetic engineering of mouse embryonic stem cells by Nurr1 enhances differentiation and maturation into dopaminergic neurons. Eur. J. Neurosci. 16 (2002) 1829–1838.

    PubMed  Article  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Kuo-Liang Yang.

Electronic supplementary material

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Yang, KL., Lee, JT., Pang, CY. et al. Human adipose-derived stem cells for the treatment of intracerebral hemorrhage in rats via femoral intravenous injection. Cell Mol Biol Lett 17, 376–392 (2012). https://doi.org/10.2478/s11658-012-0016-5

Download citation

Key words

  • Adipose-derived stem cells
  • Adult stem cells
  • Adipose tissue
  • Intracerebral hemorrhage
  • Intravenous stem cell injection
  • Neural stem cells
  • Regenerative medicine
  • Stem cell therapy
  • Stroke
  • Stromal cells