Therapeutic Impact of Human Bone Marrow Stromal Cells Expanded by Animal Serum–Free Medium for Cerebral Infarct in Rats

BACKGROUND:The donor cell culture in animal serum-free medium is important for the clinical application of cell transplantation therapy. Recently, human-derived platelet lysate (PL) gained interest as a substitute for fetal calf serum (FCS), but there are no studies that evaluate the validity of human bone marrow stromal cells (hBMSCs) expanded with PL-containing medium for central nervous system disorders. OBJECTIVE:To test the hypothesis that hBMSCs expanded with FCS-free, PL-containing medium can promote functional recovery after cerebral infarct. METHODS:hBMSCs were cultured in the FCS- or PL-containing medium. Cell-growth kinetics were analyzed. The vehicle or hBMSCs was stereotactically transplanted into the ipsilateral striatum of the rats subjected to permanent middle cerebral artery occlusion 7 days after the insult. Motor function was assessed for 8 weeks, and the fate of transplanted hBMSCs was examined using immunohistochemistry. RESULTS:There was no significant difference in hBMSC expansion between the 2 groups. Transplantation of hBMSCs expanded with the FCS- or PL-containing medium equally promoted functional recovery compared with the vehicle group. Histological analysis revealed that there were no significant differences in their migration, survival, and neural differentiation in the infarct brain between the 2 groups. CONCLUSION:hBMSCs expanded with PL-containing medium retained their capacity of migration, survival, and differentiation and significantly promoted functional recovery when stereotactically transplanted into the infarct brain. The PL may be a clinically valuable and safe substitute for FCS in expanding hBMSCs to regenerate the infarct brain.

[1]  千葉 泰弘.  Transplanted bone marrow stromal cells promote axonal regeneration and improve motor function in a rat spinal cord injury model , 2010 .

[2]  L. Pitts,et al.  Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination. , 1986, Stroke.

[3]  D. Prockop,et al.  Engraftment and migration of human bone marrow stromal cells implanted in the brains of albino rats--similarities to astrocyte grafts. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[4]  S. Kuroda,et al.  Role of SDF-1/CXCR4 system in survival and migration of bone marrow stromal cells after transplantation into mice cerebral infarct , 2007, Brain Research.

[5]  Wolfgang Wagner,et al.  Replicative senescence-associated gene expression changes in mesenchymal stromal cells are similar under different culture conditions , 2010, Haematologica.

[6]  E. Andreu,et al.  Comparison of ex vivo expansion culture conditions of mesenchymal stem cells for human cell therapy , 2009, Transfusion.

[7]  R. Walker,et al.  Development of antibodies to fetal calf serum with arthus-like reactions in human immunodeficiency virus-infected patients given syngeneic lymphocyte infusions. , 1997, Blood.

[8]  F. Prósper,et al.  Autologous human serum for cell culture avoids the implantation of cardioverter-defibrillators in cellular cardiomyoplasty. , 2004, International journal of cardiology.

[9]  A. Björklund,et al.  Immortalized neural progenitor cells for CNS gene transfer and repair , 1997, Trends in Neurosciences.

[10]  R A Swanson,et al.  A Semiautomated Method for Measuring Brain Infarct Volume , 1990, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[11]  D. Stein,et al.  Progesterone inhibits ischemic brain injury in a rat model of permanent middle cerebral artery occlusion. , 2007, Restorative neurology and neuroscience.

[12]  Qi Zhang,et al.  A combined procedure to deliver autologous mesenchymal stromal cells to patients with traumatic brain injury. , 2008, Cytotherapy.

[13]  S. Kuroda,et al.  Synergistic effects of granulocyte-colony stimulating factor on bone marrow stromal cell transplantation for mice cerebral infarct. , 2009, Cytokine.

[14]  H. Shiba,et al.  Human autologous serum obtained using a completely closed bag system as a substitute for foetal calf serum in human mesenchymal stem cell cultures , 2006, Cell biology international.

[15]  M. Chopp,et al.  Therapeutic Benefit of Intravenous Administration of Bone Marrow Stromal Cells After Cerebral Ischemia in Rats , 2001, Stroke.

[16]  Andrej-Nikolai Spiess,et al.  Accelerated and safe expansion of human mesenchymal stromal cells in animal serum‐free medium for transplantation and regenerative medicine , 2007, Journal of cellular physiology.

[17]  C. Choong,et al.  PDGF, TGF-beta, and FGF signaling is important for differentiation and growth of mesenchymal stem cells (MSCs): transcriptional profiling can identify markers and signaling pathways important in differentiation of MSCs into adipogenic, chondrogenic, and osteogenic lineages. , 2008, Blood.

[18]  A. Zander,et al.  Autologous serum for isolation and expansion of human mesenchymal stem cells for clinical use. , 2004, Experimental hematology.

[19]  H. Higuchi,et al.  Motility and growth of human bone-marrow mesenchymal stem cells during ex vivo expansion in autologous serum. , 2005, The Journal of bone and joint surgery. British volume.

[20]  Katrine Frønsdal,et al.  In Vitro Expansion of Human Mesenchymal Stem Cells: Choice of Serum Is a Determinant of Cell Proliferation, Differentiation, Gene Expression, and Transcriptome Stability , 2005, Stem cells.

[21]  C. Ide,et al.  Spinal cord injury treatment with intrathecal autologous bone marrow stromal cell transplantation: the first clinical trial case report. , 2008, The Journal of trauma.

[22]  M. Gunetti,et al.  Mesenchymal stem cell transplantation in amyotrophic lateral sclerosis: A Phase I clinical trial , 2010, Experimental Neurology.

[23]  E. Hogan,et al.  A model of focal ischemic stroke in the rat: reproducible extensive cortical infarction. , 1986, Stroke.

[24]  X. Holy,et al.  Platelet lysates promote mesenchymal stem cell expansion: A safety substitute for animal serum in cell‐based therapy applications , 2005, Journal of cellular physiology.

[25]  K. Mizuseki,et al.  Directed differentiation of telencephalic precursors from embryonic stem cells , 2005, Nature Neuroscience.

[26]  Mamoru Tamura,et al.  In vivo tracking of bone marrow stromal cells transplanted into mice cerebral infarct by fluorescence optical imaging. , 2004, Brain research. Brain research protocols.

[27]  G. Tseng,et al.  Transplanted bone marrow stromal cells migrate, differentiate and improve motor function in rats with experimentally induced cerebral stroke , 2008, Journal of anatomy.

[28]  W. Janssen,et al.  Adult Bone Marrow Stromal Cells Differentiate into Neural Cells in Vitro , 2000, Experimental Neurology.

[29]  A. Buchan,et al.  Transplantation of cultured human neuronal cells for patients with stroke , 2001, Neurology.

[30]  B. Eppley,et al.  Platelet quantification and growth factor analysis from platelet-rich plasma: implications for wound healing. , 2004, Plastic and reconstructive surgery.

[31]  Karl Kashofer,et al.  Human platelet lysate can replace fetal bovine serum for clinical‐scale expansion of functional mesenchymal stromal cells , 2007, Transfusion.

[32]  S. Kuroda,et al.  Transplanted bone marrow stromal cells improves cognitive dysfunction due to diffuse axonal injury in rats , 2009, Neuropathology : official journal of the Japanese Society of Neuropathology.

[33]  Benjamin E. Reubinoff,et al.  Neural progenitors from human embryonic stem cells , 2001, Nature Biotechnology.

[34]  R. McKay,et al.  In vitro-generated neural precursors participate in mammalian brain development. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[35]  Taku Sugiyama,et al.  Near-Infrared Fluorescence Labeling Allows Noninvasive Tracking of Bone Marrow Stromal Cells Transplanted Into Rat Infarct Brain , 2011, Neurosurgery.

[36]  M. Chopp,et al.  Therapeutic benefit of intracerebral transplantation of bone marrow stromal cells after cerebral ischemia in rats , 2001, Journal of the Neurological Sciences.

[37]  A. Cometa,et al.  Optimization of in vitro expansion of human multipotent mesenchymal stromal cells for cell‐therapy approaches: Further insights in the search for a fetal calf serum substitute , 2007, Journal of cellular physiology.

[38]  Ross Zafonte,et al.  Neurotransplantation for patients with subcortical motor stroke: a phase 2 randomized trial. , 2005, Journal of neurosurgery.

[39]  Satoshi Kuroda,et al.  Do bone marrow stromal cells proliferate after transplantation into mice cerebral infarct?—A double labeling study , 2005, Brain Research.

[40]  A. Reinisch,et al.  Platelet-derived growth factors for GMP-compliant propagation of mesenchymal stromal cells. , 2009, Bio-medical materials and engineering.

[41]  S. Totey,et al.  Ex vivo-expanded autologous bone marrow-derived mesenchymal stromal cells in human spinal cord injury/paraplegia: a pilot clinical study. , 2009, Cytotherapy.

[42]  A. Reinisch,et al.  Rapid large-scale expansion of functional mesenchymal stem cells from unmanipulated bone marrow without animal serum. , 2008, Tissue engineering. Part C, Methods.

[43]  J. Shumsky,et al.  Treatments for , 2004 .

[44]  I. Black,et al.  Adult rat and human bone marrow stromal cells differentiate into neurons , 2000, Journal of neuroscience research.

[45]  S. Kuroda,et al.  Noninvasive Transplantation of Bone Marrow Stromal Cells for Ischemic Stroke: Preliminary Study With a Thermoreversible Gelation Polymer Hydrogel , 2010, Neurosurgery.

[46]  M. Chopp,et al.  Human marrow stromal cell therapy for stroke in rat: Neurotrophins and functional recovery , 2002, Neurology.

[47]  Satoshi Kuroda,et al.  Bone marrow stromal cells protect and repair damaged neurons through multiple mechanisms , 2008, Journal of neuroscience research.

[48]  Oh Young Bang,et al.  Autologous mesenchymal stem cell transplantation in stroke patients , 2005, Annals of neurology.

[49]  A. Kiani,et al.  Treatment of refractory acute GVHD with third-party MSC expanded in platelet lysate-containing medium , 2009, Bone Marrow Transplantation.