Cell Transplantation 25(4) Abstracts

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Cell Transplantation, Vol. 25, pp. 621-628, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690872
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved


Review

Acupuncture-Induced Analgesia: The Role of Microglial Inhibition

Lili Lin,*† Nikola Skakavac,‡ Xiaoyang Lin,‡ Dong Lin,* Mia C. Borlongan,† Cesar V. Borlongan,† and Chuanhai Cao‡

*College of Acupuncture, Fujian University of Traditional Chinese Medicine, Minhou Shangjie, Fuzhou, Fujian, P. R. China
†Department of Neurosurgery and Brain Repair, University of South Florida, Morsani College of Medicine, Tampa, FL, USA
‡College of Pharmacy, Department of Pharmaceutical Sciences, University of South Florida, Morsani College of Medicine, Tampa, FL, USA

The last three decades have documented preclinical and clinical data supporting the use of acupuncture in relieving symptoms of many diseases, including allergies, infections, and neurological disorders. The advent of electroacupuncture has not only modernized the practice of acupuncture, but also has improved its efficacy, especially for producing analgesic-like effects. Although the mechanism of action of acupuncture-induced analgesia remains largely unknown, several lines of investigation have implicated modulation of pain processes via brain opioid signaling andneuroimmunoregulatory pathways. Here, we review key findings demonstrating the efficacy and underlying mechanisms of acupuncture-induced analgesia. In particular, we discuss potent analgesic effects of acupuncture via neural pain processes through inhibition of microglial activation. The safe and effective use of acupuncture stands as a nonpharmacological alternative for induction of analgesia, which has direct clinical applications, especially for pain-related diseases.

Key words: Alternative medicine; Analgesia; Inflammation; Pain; Drug-free

Received November 23, 2015; final acceptance January 29, 2016. Online prepub date: February 4, 2016.
Address correspondence to Prof. Cesar V. Borlongan, Department of Neurosurgery and Brain Repair, University of South Florida, Morsani College of Medicine, 12901 Bruce B. Downs Blvd., Tampa, FL 33612, USA. Tel: +1-813-0974-3154; Fax: +1-813-974-3078; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Dr. Chuanhai Cao, College of Pharmacy, Department of Pharmaceutical Sciences, University of South Florida, Morsani College of Medicine, 12901 Bruce B. Downs Blvd., Tampa, FL 33612, USA. Tel: +1-813-0974-3154; Fax: +1-813-974-3078; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 629-643, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690782
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Analgesic Effect of Recombinant GABAergic Cells in a Model of Peripheral Neuropathic Pain

Stanislava JergovaShyam Gajavelli, Mathew S. Varghese, Paul Shekane, and Jacqueline Sagen

Miller School of Medicine, Miami Project, University of Miami, Miami, FL, USA

Chronic neuropathic pain represents a clinically challenging state with a poor response to current treatment options. Long-term management of chronic pain is often associated with the development of tolerance, addiction, and other side effects, reducing the therapeutic value of treatment. Alternative strategies based on cell therapy and gene manipulation, balancing the inhibitory and excitatory events in the spinal cord, may provide sustained pain relief in the long term. Transplantation of GABAergic cells has been successfully used to enhance inhibition and to restore physiological spinal pain processing. However, since the underlying mechanism of chronic pain development involves changes in several pain-signaling pathways, it is essential to develop an approach that targets several components of pain signaling. Recombinant cell therapy offers the possibility to deliver additional analgesic substances to the restricted area in the nervous system. The current study explores the analgesic potential of genetically modified rat embryonic GABAergic cells releasing a peptidergic NMDA receptor antagonist, Serine1-histogranin (SHG). Overactivation of glutamate NMDA receptors contributes to the hyperexcitability of spinal neurons observed in chronic pain models. Our approach allows us to simultaneously target spinal hyperexcitability and reduced inhibitory processes. Transplantable cells were transduced by viral vectors encoding either one or six copies of SHG cDNAs. The analgesic potential of recombinant cells after their intraspinal transplantation was evaluated in a model of peripheral nerve injury. Enhanced reduction of hypersensitivity to thermal and mechanical stimuli was observed in animals treated by recombinant cells compared to the nonrecombinant group. The recombinant peptide was detected in the spinal tissue, suggesting its successful production by transplanted cells. Our results demonstrate the feasibility of using recombinant cells releasing adjunct analgesic peptides in the therapy of neuropathic pain.

Key words: GABA; Neuronal progenitor cells; Serine1-histogranin (SHG); Recombinant cells; Chronic pain

Received October 2, 2015; final acceptance January 28, 2016. Online prepub date: January 26, 2016.
Address correspondence to Stanislava Jergova, Ph.D., Miller School of Medicine, Miami Project, University of Miami, 1095 NW 14th Terrace, Miami, FL 33136, USA. Tel: +1-305-243-6038; Fax: +1-305-243-3923; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 645-664, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X690350
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Intracerebral Cell Implantation: Preparation and Characterization of Cell Suspensions

Tiziana Rossetti,* Francesca Nicholls,*† and Michel Modo*

*Departments of Radiology and Bioengineering, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA

†Institute of Psychiatry, Psychology and Neuroscience, King’s College London, London, UK

Intracerebral cell transplantation is increasingly finding a clinical translation. However, the number of cells surviving after implantation is low (5–10%) compared to the number of cells injected. Although significant efforts have been made with regard to the investigation of apoptosis of cells after implantation, very little optimization of cell preparation and administration has been undertaken. Moreover, there is a general neglect of the biophysical aspects of cell injection. Cell transplantation can only be an efficient therapeutic approach if an optimal transfer of cells from the dish to the brain can be ensured. We therefore focused on the in vitro aspects of cell preparation of a clinical-grade human neural stem cell (NSC) line for intracerebral cell implantation. NSCs were suspended in five different vehicles: phosphate-buffered saline (PBS), Dulbecco’s modified Eagle medium (DMEM), artificial cerebral spinal fluid (aCSF), HypoThermosol, andPluronic. Suspension accuracy, consistency, and cell settling were determined for different cell volume fractions in addition to cell viability, cell membrane damage, and clumping. Maintenance of cells in suspension was evaluated while being stored for 8 h on ice, at room temperature, or physiological normothermia. Significant differences between suspension vehicles and cellular volume fractions were evident. HypoThermosol and Pluronic performed best, with PBS, aCSF, and DMEM exhibiting less consistency, especially in maintaining a suspension and preserving viability under different storage conditions. These results provide the basis to further investigate these preparation parameters during the intracerebral delivery of NSCs to provide an optimized delivery process that can ensure an efficient clinical translation.

Key words: Cell transplantation; Cell suspension; Brain; Sedimentation; Volume fraction; Vehicle; Sol; Dispersed phase; Dispersion medium; HypoThermosolPluronic F68; Phosphate-buffered saline (PBS); Artificial cerebral spinal fluid (aCSF); Temperature

Received September 15, 2015; final acceptance January 29, 2016. Online prepub date: December 20, 2015.
Address correspondence to Dr. Mike Modo, McGowan Institute for Regenerative Medicine, University of Pittsburgh, 3025 East Carson St., Pittsburgh, PA 15203, USA. Tel: +1-412-383-7200; Fax: +1-412-383-6799; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 665-675, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X690297
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Direct Comparison of Rat- and Human-Derived Ganglionic Eminence Tissue Grafts on Motor Function

Mariah J. Lelos,* Victoria H. Roberton,* Ngoc-Nga Vinh,* Carl Harrison,* Peter Eriksen,* Eduardo M. Torres,* Susanne P. Clinch,* Anne E. Rosser,*† and Stephen B. Dunnett*

*Brain Repair Group, School of Biosciences, Cardiff University, Cardiff, Wales, UK
†Neuroscience and Mental Health Institute, School of Medicine, Cardiff University, Cardiff, Wales, UK

Huntington’s disease (HD) is a debilitating, genetically inherited neurodegenerative disorder that results in early loss of medium spiny neurons from the striatum and subsequent degeneration of cortical and other subcortical brain regions. Behavioral changes manifest as a range of motor, cognitive, and neuropsychiatric impairments. It has been established that replacement of the degenerated medium spiny neurons with rat-derived fetal whole ganglionic eminence (rWGE) tissue can alleviate motor and cognitive deficits in preclinical rodent models of HD. However, clinical application of this cell replacement therapy requires the use of human-derived (hWGE), not rWGE, tissue. Despite this, little is currently known about the functional efficacy of hWGE. The aim of this study was to directly compare the ability of the gold standard rWGE grafts, against the clinically relevant hWGE grafts, on a range of behavioral tests of motor function. Lister hooded rats either remained as unoperated controls or received unilateral excitotoxic lesions of the lateral neostriatum. Subsets of lesioned rats then received transplants of either rWGE or hWGEprimary fetal tissue into the lateral striatum. All rats were tested postlesion and postgraft on the following tests of motor function: staircase test, apomorphine-induced rotation, cylinder test, adjusting steps test, and vibrissae-evoked touch test. At 21 weeks postgraft, brain tissue was taken for histological analysis. The results revealed comparable improvements in apomorphine-induced rotational bias and the vibrissae test, despite larger graft volumes in the hWGE cohort. hWGE grafts, but not rWGE grafts, stabilized behavioral performance on the adjusting steps test. These results have implications for clinical application of cell replacement therapies, as well as providing a foundation for the development of stem cell-derived cell therapy products.

Key words: Huntington’s disease (HD); Motor deficits; Cell transplantation; Ganglionic eminence

Received September 25, 2015; final acceptance January 28, 2016. Online prepub date: December 29, 2015.
Address correspondence to Dr. Mariah J. Lelos, School of Biosciences, Cardiff University, Museum Avenue, Cardiff CF10 3AX, Wales, UK. Tel: +44 (0)2920 874112; Fax: +44 (0)2920 876749; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 677-686, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690863
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Allele-Specific Reduction of the Mutant Huntingtin Allele Using Transcription Activator-Like Effectors in Human Huntington’s Disease Fibroblasts

Kyle D. Fink,* Peter Deng,*† Josh Gutierrez,* Joseph S. Anderson,* Audrey Torrest,* Anvita Komarla,*† Stefanos Kalomoiris,* Whitney Cary,* Johnathon D. Anderson,* William Gruenloh,* Alexandra Duffy,‡ Teresa Tempkin,‡ Geralyn Annett,* Vicki Wheelock,‡ David J. Segal,† and Jan A. Nolta*

*Stem Cell Program and Institute for Regenerative Cures, University of California Davis Health Systems, Sacramento, CA, USA
†Genome Center, MIND Institute, and Biochemistry and Molecular Medicine, University of California, Davis, CA, USA
‡Department of Neurology, University of California Davis Health Systems, Sacramento, CA, USA

Huntington’s disease (HD) is an autosomal dominant neurodegenerative disorder caused by an abnormal expansion of CAG repeats. Although pathogenesis has been attributed to thispolyglutamine expansion, the underlying mechanisms through which the huntingtin protein functions have yet to be elucidated. It has been suggested that postnatal reduction of mutant huntingtin through protein interference or conditional gene knockout could prove to be an effective therapy for patients suffering from HD. For allele-specific targeting, transcription activator-like effectors (TALE) were designed to target single-nucleotide polymorphisms (SNP) in the mutant allele and packaged into a vector backbone containing KRAB to promote transcriptional repression of the disease-associated allele. Additional TALEs were packaged into a vector backbone containing heterodimeric FokI and were designed to be used as nucleases (TALEN) to cause a CAG-collapse in the mutant allele. Human HD fibroblasts were treated with each TALE-SNP or TALEN. Allele-expression was measured using SNP-genotyping assay and mutant protein aggregation was quantified with Western blots for anti-ubiquitin. The TALE-SNP and TALEN significantly reduced mutant allele expression (p < 0.05) when compared to control transfections while not affecting expression of the nondisease allele. This study demonstrates the potential of allele-specific gene modification using TALE proteins, and provides a foundation for targeted treatment for individuals suffering from Huntington’s or other genetically linked diseases.

Key words: Huntington’s disease (HD); Transcription activator-like effector (TALE); Gene therapy; Allele-specific silencing

Received October 31, 2015; final acceptance January 28, 2016. Online prepub date: February 4, 2016.
Address correspondence to Kyle Fink, Ph.D., Institute for Regenerative Cures, Rm 1300, 2921 Stockton Blvd., Sacramento, CA 95817, USA. Fax: +1-916-703-9310; Tel: +1-916-703-9300; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 687-703, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690818
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Mesenchymal Stem Cells as Treatment for Behavioral Deficits and Neuropathology in the 5xFAD Mouse Model of Alzheimer’s Disease

Jessica J. Matchynski-Franks,*†‡ Colleen Pappas,*† Julien Rossignol,*†§ Tiffany Reinke,*† Kyle Fink,*† Andrew Crane,*† Alison Twite,*† Steven A. Lowrance,*† Cheng Song,*† and Gary L. Dunbar*†¶#

*Field Neurosciences Laboratory for Restorative Neurology, Central Michigan University, Mt. Pleasant, MI, USA
†Program in Neuroscience, Central Michigan University, Mt. Pleasant, MI, USA
‡Department of Psychology and Behavioral Sciences, Rochester College, Rochester Hills, MI, USA
§College of Medicine, Central Michigan University, Mt. Pleasant, MI, USA
¶Department of Psychology, Central Michigan University, Mt. Pleasant, MI, USA
#St. Mary’s of Michigan Field Neurosciences Institute, Saginaw MI, USA

Alzheimer’s disease (AD) is characterized by a progressive loss of memory and other cognitive disturbances. The neuropathology of AD includes the major hallmarks of toxic amyloid-b oligomer accumulation and neurofibrillary tangles, as well as increased oxidative stress, cholinergic dysfunction, synapse loss, changes in endogenous neurotrophic factors, and overall degeneration of the brain. Adult mesenchymal stem cells (MSCs) offer the potential for a readily available treatment that would be long lasting, have low likelihood of rejection, and could target a variety of pathological deficits. MSCs have been shown to be effective in alleviating symptoms in some transgenic models of AD, but the optimal location for transplanting MSCs has yet to be determined. In the present study, the behavioral effects of transplantation of MSCs into the lateral ventricles, the hippocampus, or both of these regions were compared in the 5xFAD mouse model of AD. The results indicate that MSC transplants effectively reduce learning deficits in the 5xFAD mouse model and demonstrate a clear impact of MSCs on the levels of Ab42 in the brains of 5xFAD mice. Overall, these findings support the hypothesis that MSCs may be a viable treatment for AD, especially when injected into the lateral ventricles.

Keywords: Alzheimer’s disease (AD); Amyloid-b protein precursor; Inflammation Mesenchymal stem cells (MSCs)

Received October 16, 2015; final acceptance February 2, 2016. Online prepub date: February 2, 2016.
Address correspondence to Gary L. Dunbar, Ph.D., Health Professions Building, HP 2182, Central Michigan University, 1280 E Campus Dr., Mt. Pleasant, MI 48859, USA. Tel: +1-989-774-3282; Fax: +1-989-774-2553; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 705-714, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690557
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Effects of Aging on Neural Stem/Progenitor Cells and Oligodendrocyte Precursor Cells After Focal Cerebral Ischemia in Spontaneously Hypertensive Rats

Anna C. Liang,*1 Emiri T. Mandeville,*1 Takakuni Maki,* Akihiro Shindo,* Angel T. Som,* Naohiro Egawa,* Kanako Itoh,* Tsu Tshen Chuang,† John D. McNeish,† Julie C. Holder,‡ Josephine Lok,* Eng H. Lo,* and Ken Arai*

*Neuroprotection Research Laboratory, Departments of Radiology and Neurology, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
†Regenerative Medicine Discovery Performance Unit, GlaxoSmithKline, Boston, MA, USA
‡Regenerative Medicine Discovery Performance Unit, GlaxoSmithKline, Stevenage, UK

Aging and vascular comorbidities such as hypertension comprise critical cofactors that influence how the brain responds to stroke. Ischemic stress induces neurogenesis and oligodendrogenesisin younger brains. However, it remains unclear whether these compensatory mechanisms can be maintained even under pathologically hypertensive and aged states. To clarify the age-related remodeling capacity after stroke under hypertensive conditions, we assessed infarct volume, behavioral outcomes, and surrogate markers of neurogenesis and oligodendrogenesis in acute and subacute phases after transient focal cerebral ischemia in 3- and 12-month-old spontaneously hypertensive rats (SHRs). Hematoxylin and eosin staining showed that 3- and 12-month-old SHRs exhibited similar infarction volumes at both 3 and 14 days after focal cerebral ischemia. However, recovery of behavioral deficits (neurological score assessment and adhesive removal test) was significantly less in 12-month-old SHRs compared to 3-month-old SHRs. Concomitantly, numbers of nestin+ neural stem/progenitor cells (NSPCs) near the infarct border area or subventricularzone in 12-month-old SHRs were lower than 3-month-old SHRs at day 3. Similarly, numbers of PDGFR-α+ oligodendrocyte precursor cells (OPCs) in the corpus callosum were lower in 12-month-old SHRs at day 3. Lower levels of NSPC and OPC numbers were accompanied by lower expression levels of phosphorylated CREB. By day 14 postischemia, NSPC and OPC numbers in 12-month-old SHRs recovered to similar levels as in 3-month-old SHRs, but the numbers of proliferating NSPCs (Ki-67+nestin+ cells) and proliferating OPCs (Ki-67+PDGFR-α+ cells) remained lower in the older brains even at day 14. Taken together, these findings suggest that aging may also decrease poststroke compensatory responses for neurogenesis and oligodendrogenesis even under hypertensive conditions.

Key words: Neural stem cells; Oligodendrocyte precursor cells (OPCs); Middle cerebral artery occlusion; Stroke; Spontaneously hypertensive rat (SHR); Aging

Received November 18, 2015; final acceptance January 28, 2016. Online prepub date: January 22, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Ken Arai, Neuroprotection Research Laboratory, 13th Street, MGH East 149-2401, Charlestown, MA 02129, USA. Tel: +1-617-724-9503; Fax: +1-617-726-7830; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Eng H. Lo, Neuroprotection Research Laboratory, 13th Street, MGH East 149-2401, Charlestown, MA 02129, USA. Tel: +1-617-726-4043; Fax: +1-617-726-7830; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 715-727, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X690369
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Induction of Endothelial Phenotype From Wharton’s Jelly-Derived MSCs and Comparison of Their Vasoprotective and Neuroprotective Potential With Primary WJ-MSCs in CA1 Hippocampal Region Ex Vivo

Patrycja ObtulowiczWioletta Lech, Lukasz Strojek, Anna Sarnowska, and Krystyna Domanska-Janik

Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland

Ischemic stroke results in violent impairment of tissue homeostasis leading to severe perturbation within the neurovascular unit (NVU) during the recovery period. The aim of this study was to assess the potential of mesenchymal stem cells (MSCs) originating from Wharton’s jelly (WJ) to differentiate into functionally competent cells of endothelial lineage (WJ-EPCs). The protective effect(s) of either primary WJ-MSCs or induced WJ-EPCs was investigated and compared after oxygen–glucose deprivation (OGD) of hippocampal organotypic slices (OHC) in the indirectcoculture model. WJ-MSCs, primed in EGM-2 (Lonza commercial medium) under 5% O2, acquired cobblestone endothelial-like morphology, formed capillary-like structures and actively took up DiI-Ac-LDL. Both cell types (WJ-MSCs and WJ-EPCs) were positive for CD73, CD90, CD105, VEGFR-2, and VEGF, but only endothelial-like culture expressed vWF and PECAM-1 markers at significant levels. In the presence of either WJ-MSCs or WJ-EPCs in the compartment below OGD-injured slices, cell death and vascular atrophy in the hypoxia-sensitive CA1 region were substantially decreased. This suggests that a paracrine mechanism may mediate WJ-MSC- and WJ-EPC-dependent protection. Thus, finally, we estimated secretion of the neuro/angio/immunomodulatory molecules IL-6, TGF-β1, and VEGF by these cell cultures. We have found that release of TGF-β1 and IL-6 was TLR ligand [LPS and Poly(I:C)] concentration dependent and stronger in WJ-EPC than WJ-MSC cultures. Simultaneously, the uneven pattern of TLR receptors and modulatory cytokine gene expression was confirmed also on qRT-PCR level, but no significant differences were noticed between WJ-EPC and primary WJ-MSC cultures.

Key words: Neuroprotection; Angioprotection; Hippocampal organotypic slice culture; Wharton’s jelly epithelial progenitor cells (EPCs) and mesenchymal stem cells (MSCs); Oxygen–glucose deprivation injury

Received September 30, 2015; final acceptance January 28, 2016. Online prepub date: December 31, 2015.
Address correspondence to Krystyna Domanska-JanikMossakowski Medical Research Centre, Polish Academy of Sciences, 5 Pawinskiego Street, 02-106 Warsaw, Poland. Tel: +48 22 608 66 04; Fax: +48 22 668 55 32; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 729-734, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690791
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Fibrinogen Reduction and Motor Function Improvement by Hematopoietic Growth Factor Treatment in Chronic Stroke in Aged Mice: A Treatment Frequency Study

Yanying Liu,*1 Maria Popescu,*†1 Sharon Longo,*1 Mei Gao,* Dongliang Wang,‡ Sandra McGillis,* and Li-Ru Zhao*

*Department of Neurosurgery, State University of New York Upstate Medical University, Syracuse, NY, USA
†Department of Neuroscience and Physiology, State University of New York Upstate Medical University, Syracuse, NY, USA
‡Department of Public Health and Preventive Medicine, State University of New York Upstate Medical University, Syracuse, NY, USA

Stroke is a serious medical condition that causes long-term neurological disability in mainly elderly adults worldwide. Lack of therapy to improve functional recovery in the chronic phase of stroke is a major challenge for stroke research. Combining two hematopoietic growth factors, stem cell factor (SCF) and granulocyte-colony stimulating factor (G-CSF), our previous studies have demonstrated the neurovascular restorative efficacy of this treatment in the chronic phase of experimental stroke. Elevated plasma fibrinogen has been thought to serve as a predictor for ischemic stroke. Here we have determined the treatment frequency in reducing plasma fibrinogen and in restoring motor function in aged mice with chronic stroke. Our findings show that SCF + G-CSF treatment in chronic stroke decreases plasma fibrinogen and improves motor function in aged mice. No differences have been found between a 2-week treatment regimen and 7-day treatment in the plasma fibrinogen assay, while the 7-day treatment regimen displays a better recovery pattern with regard to motor function. This study provides new insight into understanding the potential contribution of SCF + G-CSF in both reducing the risk of recurrent ischemic stroke and enhancing stroke recovery.

Key words: Stem cell factor (SCF); Granulocyte-colony stimulating factor (G-CSF); Hematopoietic growth factor; Chronic stroke; Stroke recovery; Fibrinogen

Received November 17, 2015; final acceptance January 28, 2016. Online prepub date: January 26, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Li-Ru Zhao, M.D., Ph.D., Department of Neurosurgery, State University of New York Upstate Medical University, 750 E. Adams Street, Syracuse, NY 13210, USA. Tel: +1-315-464 8470; Fax: +1-315-464-5504; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 735-747, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690458
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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The Cytoprotective Effects of Human Endothelial Progenitor Cell-Conditioned Medium Against an Ischemic Insult Are Not Dependent on VEGF and IL-8

Stefano Di Santo, Anna-Lena Fuchs, Ramesh Periasamy, Stefanie Seiler, and Hans Rudolf Widmer

Department of Neurosurgery, Neurocenter and Regenerative Neuroscience Cluster, University of Bern, Inselspital, Bern, Switzerland

Endothelial progenitor cells (EPCs) promote revascularization and tissue repair mainly by paracrine actions. In the present study, we investigated whether EPC-secreted factors in the form of conditioned medium (EPC-CM) can protect cultured brain microvascular endothelial cells against an ischemic insult. Furthermore, we addressed the type of factors that are involved in the EPC-CM-mediated functions. For that purpose, rat brain-derived endothelial cells (rBCEC4 cell line) were exposed to EPC-CM pretreated with proteolytic digestion, heat inactivation, and lipid extraction. Moreover, the involvement of VEGF and IL-8, as canonical angiogenic factors, was investigated by means of neutralizing antibodies. We demonstrated that EPC-CM significantly protected the rBCEC4 cells against an ischemic insult mimicked by induced oxygen–glucose deprivation followed by reoxygenation. The cytoprotective effect was displayed by higher viable cell numbers and reduced caspase 3/7 activity. Heat inactivation, proteolytic digestion, and lipid extraction resulted in a significantly reduced EPC-CM-dependent increase in rBCEC4 viability, tube formation, and survival following the ischemic challenge. Notably, VEGF and IL-8 neutralization did not affect the actions of EPC-CM on rBCEC4 under both standard and ischemic conditions. In summary, our findings show that paracrine factors released by EPCs activate an angiogenic and cytoprotective response on brain microvascular cells and that the activity of EPC-CM relies on the concerted action of nonproteinaceous and proteinaceous factors but do not directly involve VEGF and IL-8.

Key words: Endothelial progenitor cells (EPCs); Paracrine factors; Cytoprotection; Ischemia; Angiogenesis

Received September 15, 2015; final acceptance January 28, 2016. Online prepub date: January 15, 2016.
Address correspondence to Hans R. Widmer, Ph.D., Department of Neurosurgery, Pavillon 47, University of Bern, InselspitalFreiburgstrasse, CH-3010 Bern, Switzerland. Tel: +41 31 632 2770; Fax: +41 31 382 2414; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it