Therapeutic Benefit of Intravenous Administration of Human Umbilical Cord Blood- Mononuclear Cells Following Intracerebral Hemorrhage in Rat

Document Type: Original Article


1 Department of Anatomy and Cell Biology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Department of Physiology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

3 Department of Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran


Human umbilical cord blood (HUCB) is now considered as a valuable source for stem cell–based therapies. Previous studies showed that intravascular injection of the HUCB significantly improves neurological functional recovery in a rat model of intracerebral hemorrhage (ICH). In the present study, we hypothesize transplanted HUCB derived mononuclear cells (UC-MCs) can decrease injured volume and also ameliorate neurological function in ICH rats.
Materials and Methods
Experimental ICH was induced by intrastriatal administration of collagenase in rats. One day after surgery, the rats were divided into 3 groups to receive intravenously either BrdU positive human UC-MCs [(4×106 and 8×106 cells in 1 ml saline, n=10 respectively) as treated groups] or the same amount of saline [as lesion group (n=10)]. There was also one group (control) that received only vehicle solution of collagenase. The animals were evaluated for 14 days with behavioral tests. Transplanted UC-MCs were detected by immunohistochemistry. Histological data and scores of functional tests were analyzed using ANOVA. Cellular co-localization of BrdU+ cells in the histological slides was determined by software Image J.
Intravenously transplanted UC-MCs migrated selectively to the hematomal area and reduce injured volume. The UC-MCs transplanted groups showed better performance on functional tests after 2 weeks compared with the lesion and control groups (P< 0.05). There was no difference in the functional recovery and injured volume improvement between the 2 treated groups.
Intravenously transplanted UC-MCs accelerate neurological function recovery of ICH rat and diminish the striatum lesion size. Thus these cells may provide a potential cell candidate for cell-based therapy in ICH.


1. Ribo M, Grotta JC. Latest advances in intracerebral hemorrhage. Curr Neurol Neurosci Rep 2006; 6:17-22.

2. Broderick JP, Adams Jr HP, Barsan W, Feinberg W, Feldmann E, Grotta J, et al. 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 1999; 30:905–15.

3. Foulkes MA, Wolf PA, Price TR, Mohr JP, Hier DB. The stroke data bank: design methods and baseline characteristics. Stroke 1988; 19:547–554.

4. Allahtavakoli M, Pourshanazari A, Heshmatian B. Vitamin E derivative alpha-tocotrienol failed to show neuroprotective effects after embolic Stroke in rats. Iran J Basic Med Sci 2009; 12: 9-17.

5. Jeong SW, Chu K, Jung KH, Kim SU, Kim M, Roh JK.  Human neural stem cell transplantation promotes functional recovery in rats with experimental intracerebral hemorrhage. Stroke 2003; 34:2258-2263.

6. Liao W, Zhong J, Yu J, Xie J, Liu Y, Du L, et al. Therapeutic benefit of human umbilical cord derived mesenchymal stromal cells in intracerebral hemorrhage rat: implications of anti-inflammation and angiogenesis. Cell Physiol Biochem 2009; 24:307-316.

7. Harris DT. Cord blood stem cells: A review of potential neurological applications. Stem Cell Rev 2008; 4:269-274.

8. Bliss T, Guzman R, Daadi M, Steinberg GK. Cell transplantation therapy for stroke. Stroke 2007; 38:817–826.

9. Broxmeyer HE, Douglas GW, Hangoc G, Cooper S,  Bard J, English D, et al. Human umbilical cord blood as a potential source of transplantable hematopoietic stem/progenitor cells. Proc Natl Acad Sci USA 1989; 86:3828–3832.

10. Erices A, Conget P, Minguell JJ. Mesenchymal progenitor cells in human umbilical cord blood. Br J Haematol 2000; 109:235–242.

11. Murohara T, Ikeda H, Duan J, Shintani S, Sasaki K, Eguchi H, et al. Transplanted cord blood-derived endothelial precursor cells augment postnatal neovascularization. J Clin. Invest 2000; 105:1527–1536.

12. Madlambayan G, Rogers I. Umbilical cord-derived stem cells for tissue therapy: current and future uses. Regen Med 2006; 1:777–787.

13. Vendrame M, Cassady J, Newcomb J, Butler T, Pennypacker KR,  Zigova T, et al. Infusion of human umbilical cord blood cells in a rat model of stroke dose-dependently rescues behavioral deficits and reduces infarct volume. Stroke 2004; 35:2390–2395.

14. Vendrame M, Gemma C, de Mesquita D, Collier L, Bickford PC, Sanberg CD, et al. Anti-infl ammatory effects of human cord blood cells in a rat model of stroke. Stem Cells Dev  2005; 14:595–604.

15. Taguchi A, Soma T, Tanaka H, Kanda T, Nishimura H, Yoshikawa H, et al. Administration of CD34+ cells after stroke enhances neurogenesis via angiogenesis in a mouse model. J Clin Invest 2004; 114:330–338.

16. Chen J, Sanberg PR, Li Y, Wang L, Lu M, Willing AE, et al.  Intravenous administration of human umbilical cord blood reduces behavioral deficits after stroke in rats. Stroke  2001; 32:2682–2688.

17. Sanberg PR, Willing AE, Garbuzova-Davis S, Saporta S, Liu G, Sanberg CD, et al. Umbilical cord blood-derived stem cells and brain repair". Ann N Y Acad Sci 2005; 1049:67– 83.

18. Park DH, Borlongan CV, Willing AE, Eve DJ, Cruz LE, Sanberg CD,  et al. Human umbilical cord blood cell grafts for brain ischemia. Cell Transplant  2009; 18:985–998.

19. Del Bigio MR, Yan HJ, Buist R, Peeling J. Experimental intracerebral hemorrhage in rats: magnetic resonance imaging and histopathological correlates. Stroke 1996; 27:2312-2319.

20. Lee ST, Chu K, Sinn DI, Jung KH, Kim EH, Kim SJ, et al. Erythropoietin reduces perihematomal inflammation and cell death with eNOS and STAT3 activations in experimental intracerebral hemorrhage. J Neurochem 2006; 96:728-1739.

21. Jalali M, Ghafaripoor HA, Nikravesh MR., Hamidi Alamdari D, Sanchooli J, Seghatoleslam M. Therapeutic potential of cord blood stem cell in brain damage of an animal model. J Iran Anatom Sci 2011; 9:191-202.

22. Sirchia G, Rebulla P. Placental/umbilical cord blood transplantation. Haematologica 1999; 84:738-747.

23. Pimentel-Coelho PM, Magalhães ES, Lopes LM, deAzevedo LC, Santiago MF, Mendez-Otero R. Human cord blood transplantation in a neonatal rat model of hypoxic-ischemic brain damage: functional outcome related to neuroprotection in the striatum. Stem Cells Dev  2010; 19:351-358.

24. Hassanein SM, Amer HA, Shahab AA, Hellal MM. Umbilical cord blood CD45 (+) CD34 (+) cells coexpression in preterm and full-term neonates: a pilot study. J Matern Fetal Neonatal Med 2011; 24:229-33.

25. De Ryck M, Van Reempts J, Borgers M, Wauquier A,Janssen PA. Photochemical stroke model: flunarizine prevents sensorimotor deficits after neocortical infarcts in rats. Stroke  1989; 20:1383-1390.

26. Hua Y, Schallert T, Keep RF, Wu J, Hoff JT, Xi G. Behavioral tests after intracerebral hemorrhage in the rat. Stroke  2002; 33:2478-2484.

27. Zhang L, Schallert T, Zhang ZG, Jiang Q, Arniego P, Li Q, et al. A test for detecting long-term sensorimotor dysfunction in the mouse after focal cerebral ischemia. J Neurosci Methods  2002; 117:207-214.

28. Riegelsberger UM, Deten A, Pösel C, Zille M, Kranz A, Boltze J, et al.  Intravenous human umbilical cord blood transplantation for stroke: impact on infarct volume and caspase-3-dependent cell death in spontaneously hypertensive rats. Exp Neurol 2011; 227:218-223.

29. Andres RH, Guzman R, Ducray AD, Mordasini P, Gera A, Barth A, et al. Cell replacement therapy for intracerebral hemorrhage.  Neurosurg Focus  2008; 24:E16.

30. Lee HJ, Kim KS, Kim EJ, Choi HB, Lee KH, , Park IH, et al. Brain transplantation of immortalized human neural stem cells promotes functional recovery in mouse intracerebral hemorrhage stroke model. Stem Cells  2007; 25:1204-1212.

31. Nonaka M, Yoshikawa M, Nishimura F, Yokota H, Kimura H, Hirabayashi H, et al. Intraventricular transplantation of embryonic stem cell-derived neural stem cells in intracerebral hemorrhage rats. Neurol Res  2004; 26:265-267.

32. Seyfried D, Ding J, Han Y, Li Y, Chen J, Chopp M. Effects of intravenous administration of human bone marrow stromal cells after intracerebral hemorrhage in rats. J Neurosurg  2006; 104:313-318.

33. Mahmood A, Lu D, Wang L, 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 2002; 19:1609-1617.

34. Zhang H, Huang Z, Xu Y, Zhang S. Differentiation and neurological benefit of the mesenchymal stem cells transplanted into the rat brain following intracerebral hemorrhage. Neurol Res 2006; 28:104-112.

35. Kim JM, Lee ST, Chu K, Jung KH, Song EC, Kim SJ, et al. Systemic transplantation of human adipose stem cells attenuated cerebral inflammation and degeneration in a hemorrhagic stroke model. Brain Res 2007; 1183:43-50.

36. Nan Z, Grande A, Sanberg CD, Sanberg PR, Low WC. Infusion of human umbilical cord blood ameliorates neurologic deficits in rats with hemorrhagic brain injury. Ann N Y Acad Sci  2005; 1049:84-96.

37. Liu AM, Lu G, Tsang KS, Li G, Wu Y, Huang ZS, et al. Umbilical cord-derived mesenchymal stem cells with forced expression of hepatocyte growth factor enhance remyelination and functional recovery in a rat intracerebral hemorrhage model. Neurosurgery 2010; 67:357-365.

38. Riordan NH, Chan K, Marleau AM, Ichim TE. Cord blood in regenerative medicine: do we need immune suppression? J Transl Med  2007; 5:8.

39. Bakhtiary M, Marzban M, Mehdizadeh M, Joghataei MT, Khoei S, Tondar M, et al. Combination of stem cell mobilized by granulocyte-colony stimulating factor and human umbilical cord matrix stem cell: therapy of traumatic brain injury in rats. Iran J Basic Med Sci  2011; 14:327-339.

40. Halabian R, Mohammadi MH, Salimi M, Amani M, Mohammadi Roushande A, Aghaipoor M, et al. Genetically engineered mesenchymal stem cells stably expressing green fluorescent protein. Iran J Basic Med Sci  2011; 13:24-30.

41. Newman M, Willing A, Cassady CJ, Manresa JJ, Kedziorek DA, Davis C, et al. In vitro migration and phenotype identification of human umbilical cord blood (HUCB) cells to stroke brain. Exp Neurol 2003; 181:84–112.

42. Li Y, Chen J, Chen XG, Wang L, Gautam SC, Xu YX, et al. Human marrow stromal cell therapy for stroke in rat: neurotrophins and functional recovery. Neurology 2002; 59:514–523.

43. Mahmood A, Lu D, Lu M, Chopp M. Treatment of traumatic brain injury in adult rats with intravenous administration of human bone marrow stromal cells. Neurosurgery 2003; 53:697–702.

44. Pomyje J, Zivny J, Sefc L, Plasilova M, Pytlik R, Necas E. Expression of genes regulating angiogenesis in human circulating hematopoietic cord blood CD34 /CD133 cells. Eur J Haematol 2003; 70:143–150.

45. Gong C, Hoff JT, Kee RF. Acute inflammatory reaction following experimental intracerebral hemorrhage in rat. Brain Res 2000; 871:57-65.

46. Rainsford E, Reen DJ. Interleukin 10, produced in abundance by human newborn T cells, may be the regulator of increased tolerance associated with cord blood stem cell transplantation. Br J Haematol 2002; 116:702–709.

47. Bracci-Laudiero L, Celestino D, Starace G, Antonelli A, Lambiase A, Procoli A, et al. CD34-positive cells in human umbilical cord blood express nerve growth factor and its specific receptor. J Neuroimmunol 2003; 136:130–139.

48. Borlongan CV, Hadman M, Davis Sanberg C, Sanberg PR. CNS entry of peripherally injected umbilical cord blood cells is not required for neuroprotection in stroke. Stroke 2004; 35:2385–2389.