Cell Transplantation 26(4) Abstracts

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Cell Transplantation, Vol. 26, pp. 531-539, 2017
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DOI: https://doi.org/10.3727/096368916X
693699
E-ISSN 1555-3892
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Review

Is Immune Modulation the Mechanism Underlying the Beneficial Effects of Amniotic Cells and Their Derivatives in Regenerative Medicine?

Antonietta R. Silini,* Marta Magatti,* Anna Cargnoni,* and Ornella Parolini*†

*Centro di Ricerca “E. Menni”, Fondazione Poliambulanza-Istituto Ospedaliero, Brescia, Italy
Istituto di Anatomia UmanaBiologia CellulareUniversità Cattolica del Sacro Cuore Facoltà di MedicinaChirurgia, Rome, Italy

Regenerative medicine aims to repair and regenerate damaged cells, tissues, and organs in order to restore function. Regeneration can be obtained either by cell replacement or by stimulating the body’s own repair mechanisms. Importantly, a favorable environment is required before any regenerative signal can stimulate resident stem/stromal cells, and regeneration is possible only after the resolution of injury-induced inflammation. An exacerbated immune response is often present in cases of degenerative, inflammatory-based diseases. Here we discuss how amniotic membrane cells, and their derivatives, can contribute to the resolution of many diseases with altered immune response by acting on different inflammatory mediators.

Key words: Human placenta; Amniotic membrane (AM); Amniotic mesenchymal stromal/stem cells; Amniotic epithelial cells; Immune modulation; Paracrine mechanism

Received September 28, 2016; final acceptance December 12, 2016. Online prepub date: November 3, 2016.
Address correspondence to Ornella Parolini, Ph.D., Istituto di Anatomia UmanaBiologia CellulareUniversità Cattolica del Sacro Cuore Facoltà di MedicinaChirurgia, Largo Francesco Vito, 1 00168 Rome, Italy. Tel: +39 030 351 8904; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it
or This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 541-553, 2017
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DOI: https://doi.org/10.3727/096368916X
693572
E-ISSN 1555-3892
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Human Amnion Epithelial Cells Protect Against White Matter Brain Injury After Repeated Endotoxin Exposure in the Preterm Ovine Fetus

Tamara Yawno,* Tharani Sabaretnam,* Jingang Li,* Courtney McDonald,* Rebecca Lim,* Graham Jenkin,*† Euan M. Wallace,*† and Suzanne L. Miller*†

*The Ritchie Centre, Hudson Institute of Medical Research, Monash University, Clayton, Victoria, Australia
†Department of Obstetrics and Gynaecology, Monash University, Clayton, Victoria, Australia

Intrauterine inflammation is a significant cause of injury to the developing fetal brain. Using a preterm fetal sheep model of in utero infection, we asked whether human amnion epithelial cells (hAECs) were able to reduce inflammation-induced fetal brain injury. Surgery was undertaken on pregnant sheep at ~105 days gestation (term is 147 days) for implantation of vascular catheters. Lipopolysaccharide (LPS; 150 ng/kg bolus) or saline was administered IV at 109, 110, and 111 days. Sixty million fluorescent-labeled hAECs were administered at 110, 111, and 112 days gestation via the brachial artery catheter. Brains were collected at 114 days for histological assessment. hAECs were observed within the cortex, white matter, and hippocampus. Compared to control lambs, LPS administration was associated with significant and widespread fetal brain inflammation and injury as evidenced by increased number of activated microglia in the periventricular white matter (
p = 0.02), increased pyknosis, cell degeneration ( p = 0.01), and a nonsignificant trend of fewer oligodendrocytes in the subcortical and periventricular white matter. Administration of hAECs to LPS-treated animals was associated with a significant mitigation in both inflammation and injury as evidenced by fewer activated microglia ( p = 0.03) and pyknotic cells ( p = 0.03), significantly more oligodendrocytes in the subcortical and periventricular white matter ( p = 0.01 and 0.02, respectively), and more myelin basic protein-positive cells within the periventricular white matter ( p = 0.02). hAEC administration to fetal sheep exposed to multiple doses of LPS dampens the resultant fetal inflammatory response and mitigates associated brain injury. 

Key words: Human amnion epithelial cells (hAECs); Lipopolysaccharide (LPS); Inflammation; Brain development; Oligodendrocytes; Fetal sheep

Received September 22, 2016; final acceptance December 6, 2016. Online prepub date: October 31, 2016.
Address correspondence to Tamara Yawno, The Ritchie Centre, Hudson Institute of Medical Research, 27-31 Wright Street, Clayton, Victoria 3168, Australia. Tel: +(61) 3-8572 2798; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 555-569, 2017
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DOI: https://doi.org/10.3727/096368916X
693842
E-ISSN 1555-3892
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Repetitive and Prolonged Omega-3 Fatty Acid Treatment After Traumatic Brain Injury Enhances Long-Term Tissue Restoration and Cognitive Recovery

Hongjian Pu,*† Xiaoyan Jiang,†‡ Zhishuo Wei,† Dandan Hong,† Sulaiman Hassan,† Wenting Zhang,‡ Jialin Liu,* Hengxing Meng,* Yejie Shi,† Ling Chen,* and Jun Chen†‡

*Department of Neurosurgery, General Hospital of PLA, Beijing, P.R. China
†Pittsburgh Institute of Brain Disorders and Recovery and Department of Neurology, University of Pittsburgh, Pittsburgh, PA, USA
‡State Key Laboratory of Medical Neurobiology and Institute of Brain Sciences, Fudan University, Shanghai, P.R. China

Traumatic brain injury (TBI) is one of the most disabling clinical conditions that could lead to neurocognitive disorders in survivors. Our group and others previously reported that prophylactic enrichment of dietary omega-3 polyunsaturated fatty acids (n-3 PUFAs) markedly ameliorate cognitive deficits after TBI. However, it remains unclear whether a clinically relevant therapeutic regimen with n-3 PUFAs administered after TBI would still offer significant improvement of long-term cognitive recovery. In the present study, we employed the decline of spatial cognitive function as a main outcome after TBI to investigate the therapeutic efficacy of post-TBI n-3 PUFA treatment and the underlying mechanisms. Mice were subjected to sham operation or controlled cortical impact, followed by random assignment to receive the following four treatments: (1) vehicle control; (2) daily intraperitoneal injections of n-3 PUFAs for 2 weeks, beginning 2 h after TBI; (3) fish oil dietary supplementation throughout the study, beginning 1 day after TBI; or (4) combination of treatments (2) and (3). Spatial cognitive deficits and chronic brain tissue loss, as well as endogenous brain repair processes such as neurogenesis, angiogenesis, and oligodendrogenesis, were evaluated up to 35 days after TBI. The results revealed prominent spatial cognitive deficits and massive tissue loss caused by TBI. Among all mice receiving post-TBI n-3 PUFA treatments, the combined treatment of fish oil dietary supplement and n-3 PUFA injections demonstrated a reproducible beneficial effect in attenuating cognitive deficits although without reducing gross tissue loss. Mechanistically, the combined treatment promoted post-TBI restorative processes in the brain, including generation of immature neurons, microvessels, and oligodendrocytes, each of which was significantly correlated with the improved cognitive recovery. These results indicated that repetitive and prolonged n-3 PUFA treatments after TBI are capable of enhancing brain remodeling and could be developed as a potential therapy to treat TBI victims in the clinic.

Key words: Omega-3 polyunsaturated fatty acid (n-3 PUFA); Water maze; Hippocampus; Angiogenesis; Oligodendrogenesis

Received November 24, 2016; final acceptance January 9, 2017. Online prepub date: November 24, 2016.
Address correspondence to Dr. Jun Chen, Pittsburgh Institute of Brain Disorders and Recovery, University of Pittsburgh, S507 Biomedical Science Tower, 3500 Terrace Street, Pittsburgh, PA 15213, USA. Tel: +1 (412) 648-1263; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it
or Dr. Ling Chen, Department of Neurosurgery, General Hospital of PLA, 28 Fuxing Road, Beijing 100853, P.R. China. Tel: +86 (10) 83198852; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 571-583, 2017
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DOI: https://doi.org/10.3727/096368916X
693563
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A Novel CXCR4 Antagonist CX549 Induces Neuroprotection in Stroke Brain

Kuo-Jen Wu,* Seong-Jin Yu,* Kak-Shan Shia,† Chien-Huang Wu,† Jen-Shin Song,† Hsuan-Hao Kuan,† Kai-Chia Yeh,† Chiung-Tong Chen,† Eunkyune Bae,* and Yun Wang*

*Center for Neuropsychiatric Research, National Health Research Institutes (NHRI), Miaoli, Taiwan
†Institute of Biotechnology and Pharmaceutical Research, NHRI, Miaoli, Taiwan

C-X-C chemokine receptor type 4 (CXCR4) is a receptor for a pleiotropic chemokine CXCL12. Previous studies have shown that the acute administration of the CXCR4 antagonist AMD3100 reduced neuroinflammation in stroke brain and mobilized bone marrow hematopoietic stem cells (HSCs). The purpose of this study was to characterize the neuroprotective and neurotrophic effect of a novel CXCR4 antagonist CX549. We demonstrated that CX549 had a higher affinity for CXCR4 and was more potent than AMD3100 to inhibit CXCL12-mediated chemotaxis in culture. CX549 effectively reduced the activation of microglia and improved neuronal survival after injury in neuron/microglia cocultures. Early poststroke treatment with CX549 significantly improved behavioral function, reduced brain infarction, and suppressed the expression of inflammatory markers. Compared to AMD3100, CX549 has a higher affinity for CXCR4, is more efficient to mobilize HSCs for transplantation, and induces behavioral improvement. Our data support that CX549 is a potent anti-inflammatory agent, is neuroprotective against ischemic brain injury, and may have clinical implications for the treatment of stroke.

Key words: C-X-C chemokine receptor type 4 (CXCR4); Stroke; Protection; Inflammation

Received September 9, 2016; final acceptance February 20, 2017. Online prepub date: October 27, 2016.
Address correspondence to Yun Wang, M.D., Ph.D., Center for Neuropsychiatric Research, National Health Research Institutes, 35 Keyan Road, Zhunan, Miaoli 35053, Taiwan. Tel: +886-37246166, ext. 36700; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 585-603, 2017
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DOI: https://doi.org/10.3727/096368916X
693671
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A Comparative Study of Three Different Types of Stem Cells for Treatment of Rat Spinal Cord Injury

Jiri Ruzicka,*1 Lucia Machova-Urdzikova,*1 John Gillick,† Takashi Amemori,* Nataliya Romanyuk,* Kristyna Karova,*‡ Kristyna Zaviskova,*‡ Jana Dubisova,*‡ Sarka Kubinova,* Raj Murali,† Eva Sykova,*‡ MeenaJhanwar-Uniyal,† and Pavla Jendelova*‡

*Institute of Experimental Medicine, Czech Academy of Sciences, Prague, Czech Republic
† New York Medical College, Valhalla, NY, USA
‡Department of Neuroscience, Charles University, Second Faculty of Medicine, Prague, Czech Republic

Three different sources of human stem cells—bone marrow-derived mesenchymal stem cells (BM-MSCs), neural progenitors (NPs) derived from immortalized spinal fetal cell line (SPC-01), and induced pluripotent stem cells (iPSCs)—were compared in the treatment of a balloon-induced spinal cord compression lesion in rats. One week after lesioning, the rats received either BM-MSCs (intrathecally) or NPs (SPC-01 cells or iPSCNPs, bothintraspinally), or saline. The rats were assessed for their locomotor skills (BBB, flat beam test, and rotarod). Morphometric analyses of spared white and gray matter, axonal sprouting, and glial scar formation, as well as qPCR and Luminex assay, were conducted to detect endogenous gene expression, while inflammatory cytokine levels were performed to evaluate the host tissue response to stem cell therapy. The highest locomotor recovery was observed in iPSC-NP-grafted animals, which also displayed the highest amount of preserved white and gray matter. Grafted iPSC-NPs and SPC-01 cells significantly increased the number of growth-associated protein 43 (GAP43+) axons, reduced astrogliosis, downregulated Casp3 expression, and increased IL-6 and IL-12 levels. hMSCs transiently decreased levels of inflammatory IL-2 and TNF-
a. These findings correlate with the short survival of hMSCs, while NPs survived for 2 months and matured slowly into glia- and tissue-specific neuronal precursors. SPC-01 cells differentiated more in astroglial phenotypes with a dense structure of the implant, whereas iPSC-NPs displayed a more neuronal phenotype with a loose  structure of the graft. We concluded that the BBB scores of iPSC-NP- and hMSC-injected rats were superior to the SPC-01-treated group. The iPSC-NP treatment of spinal cord injury (SCI) provided the highest recovery of locomotor function due to robust graft survival and its effect on tissue sparing, reduction of glial scarring, and increased axonal sprouting.

Key words: Spinal cord injury (SCI); iPSC-derived human neural progenitors; Inflammatory response Human fetal neural stem cells; Human mesenchymal stem cells (hMSCs);

Received September 19, 2016; final acceptance January 27, 2017. Online prepub date: November 2, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Pavla JendelovaVidenska 1083, Prague 4, Prague, Czech Republic. Tel: (+420 2) 4106 2828; Fax: (+420 2) 4106 2706; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it
or Meena Jhanwar-Uniyal, New York Medical College, Valhalla, NY, USA. Tel: 1(914)594-2513; Fax: 1(914) 594-4002; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 605-611, 2017
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DOI: https://doi.org/10.3727/096368917X
694840
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Fate of Neural Progenitor Cells Transplanted Into Jaundiced and Nonjaundiced Rat Brains

Fu-Chen Yang,* Sean M. Riordan,† Michelle Winter,‡ Li Gan,* Peter G. Smith,*‡ Jay L. Vivian,§ Steven M. Shapiro,† and John A. Stanford*‡

*Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS, USA
†Division of Child Neurology, Department of Pediatrics, Children’s Mercy Hospital and Clinics, Kansas City, MO, USA
‡Kansas Intellectual and Developmental Disabilities Research Center, University of Kansas Medical Center, Kansas City, KS, USA
§Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, KS, USA

High levels of bilirubin in infants can cause kernicterus, which includes basal ganglia damage and dystonia. Stem cell transplantation may be an effective treatment for this disease. In this study, we transplanted human neural progenitor cells differentiated toward propriospinal interneurons into the striatum of 20-day-old spontaneously jaundiced (jj) Gunn rats and nonjaundiced (Nj) littermates. Using immunohistochemical methods, we found that grafted cells survived and grew fibers in jj and Nj brains 3 weeks after transplantation. Grafted cells had a higher survival rate in jj than in Nj brains, suggesting that slightly elevated bilirubin may protect graft survival due to its antioxidative and immunosuppressive effects. Despite their survival, only a small portion of grafted neurons expressed GAD-6 or ChAT, which mark GABAergic and cholinergic neurons, respectively, and are the cells that we are attempting to replace in kernicterus. Thus, NPCs containing large populations of GABAergic and cholinergic neurons should be used for further study in this field.

Key words: Bilirubin encephalopathy; Basal ganglia; Gunn rats; Xenotransplantation

Received September 15, 2016; final acceptance January 9, 2017. Online prepub date: February 3, 2017.
Address correspondence to John A. Stanford, Ph.D., Department of Molecular and Integrative Physiology, University of Kansas Medical Center, 3901 Rainbow Boulevard, MS 3051, Kansas City, KS 66160, USA. Tel: 913-588-7416; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 613-624, 2017
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DOI: https://doi.org/10.3727/096368916X
692979
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Real-Time Intraoperative MRI Intracerebral Delivery of Induced Pluripotent Stem Cell-Derived Neurons

Scott C. Vermilyea,*†1 Jianfeng Lu,‡1 Miles Olsen,§ Scott Guthrie,* Yunlong Tao,‡ Eva M. Fekete,¶ Marissa K. Riedel,¶ Kevin Brunner,* Carissa Boettcher,* Viktorya Bondarenko,* Ethan Brodsky,§ Walter F. Block,§# Andrew Alexander,*‡§¶ Su-Chun Zhang,†** and Marina E. Emborg*†§

*Preclinical Parkinson’s Research Program, Wisconsin National Primate Research Center, University of Wisconsin, Madison, WI, USA
†Neuroscience Training Program, University of Wisconsin, Madison, WI, USA
Waisman Center, University of Wisconsin, Madison, WI, USA
§Department of Medical Physics, University of Wisconsin, Madison, WI, USA
¶Department of Psychiatry, University of Wisconsin, Madison, WI, USA
#Department of Biomedical Engineering, University of Wisconsin, Madison, WI, USA
**Department of Neuroscience, University of Wisconsin, Madison, WI, USA

Induced pluripotent stem cell (iPSC)-derived neurons represent an opportunity for cell replacement strategies for neurodegenerative disorders such as Parkinson’s disease (PD). Improvement in cell graft targeting, distribution, and density can be key for disease modification. We have previously developed a trajectory guide system for real-time intraoperative magnetic resonance imaging (RT-IMRI) delivery of infusates, such as viral vector suspensions for gene therapy strategies. Intracerebral delivery of iPSC-derived neurons presents different challenges than viral vectors, including limited cell survival if cells are kept at room temperature for prolonged periods of time, precipitation and aggregation of cells in the cannula, and obstruction during injection, which must be solved for successful application of this delivery approach. To develop procedures suitable for RT-IMRI cell delivery, we first performed in vitro studies to tailor the delivery hardware (e.g., cannula) and defined a range of parameters to be applied (e.g., maximal time span allowable between cell loading in the system and intracerebral injection) to ensure cell survival. Then we performed an in vivo study to evaluate the feasibility of applying the system to nonhuman primates. Our results demonstrate that the RT-IMRI delivery system provides valuable guidance, monitoring, and visualization during intracerebral cell delivery that are compatible with cell survival.

Key words: Magnetic resonance imaging (MRI); Induced pluripotent stem cells (iPSCs); Intracerebral delivery; Intracerebral targeting; Stereotaxic surgery; Putamen

Received November 19, 2015; final acceptance October 12, 2016. Online prepub date: September 14, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Marina E. Emborg, M.D., Ph.D., Preclinical Parkinson’s Research Program, Wisconsin National Primate Research Center, University of Wisconsin-Madison, 1223 Capitol Court, Madison, WI 53715, USA. Tel: 608-262-9714; Fax: 608-263-3524; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 625-645, 2017
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DOI: https://doi.org/10.3727/096368916X
693680
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A Comparison of Exogenous Labels for the Histological Identification of Transplanted Neural Stem Cells

Francesca J. Nicholls,*†‡ Jessie R. Liu,§ and Michel Modo*†§

*McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA
†Department of Radiology, University of Pittsburgh, Pittsburgh, PA, USA
‡Department of Neuroscience, Institute of Psychiatry, King’s College London, London, UK
§Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA, USA

The interpretation of cell transplantation experiments is often dependent on the presence of an exogenous label for the identification of implanted cells. The exogenous labels Hoechst 33342, 5-bromo-2
¢-deoxyuridine (BrdU), PKH26, and Qtracker were compared for their labeling efficiency, cellular effects, and reliability to identify a human neural stem cell (hNSC) line implanted intracerebrally into the rat brain. Hoechst 33342 (2 mg/ml) exhibited a delayed cytotoxicity that killed all cells within 7 days. This label was hence not progressed to in vivo studies. PKH26 (5 mM), Qtracker (15 nM), and BrdU (0.2 mM) labeled 100% of the cell population at day 1, although BrdU labeling declined by day 7. BrdU and Qtracker exerted effects on proliferation and differentiation. PKH26 reduced viability and proliferation at day 1, but this normalized by day 7. In an in vitro coculture assay, all labels transferred to unlabeled cells. After transplantation, the reliability of exogenous labels was assessed against the gold standard of a human-specific nuclear antigen (HNA) antibody. BrdU, PKH26, and Qtracker resulted in a very small proportion (<2%) of false positives, but a significant amount of false negatives (~30%), with little change between 1 and 7 days. Exogenous labels can therefore be reliable to identify transplanted cells without exerting major cellular effects, but validation is required. The interpretation of cell transplantation experiments should be presented in the context of the label’s limitations.

Key words: Neural stem cells (NSCs); Cell transplantation; Hoechst; PKH26; Quantum dots; Qtracker; 5-Bromo-2¢-deoxyuridine (BrdU); Validation; Proliferation; Differentiation; Toxicity; Phototoxicity; False positive; False negative; Reliability

Received September 22, 2016; final acceptance January 26, 2017. Online prepub date: November 3, 2016.
Address correspondence to Mike Modo, McGowan Institute for Regenerative Medicine and Department of Radiology, University of Pittsburgh, 3025 East Carson Street, Pittsburgh, PA 15203, USA. Tel: +1 (412) 383 7200; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 647-658, 2017
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DOI: https://doi.org/10.3727/096368916X
693716
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Transplantation of Mesenchymal Stromal Cells in Patients With Amyotrophic Lateral Sclerosis: Results of Phase I/IIa Clinical Trial

Eva Syková,*† Petr Rychmach,‡ Ivana Drahorádová,‡ Šimona Konrádová,*‡ Kateřina Růžičková,*‡ Ivan Voříšek,* Serhiy Forostyak,*† Aleš Homola,†§ and Martin Bojar§

*Department of Neuroscience, Institute of Experimental Medicine, Czech Academy of Sciences, Prague, Czech Republic
†Department of Neuroscience, Charles University, 2nd Medical School, Prague, Czech Republic
Bioinova Ltd., Prague, Czech Republic
§Department of Neurology, Charles University, 2nd Medical School, University Hospital at Motol, Prague, Czech Republic

Amyotrophic lateral sclerosis (ALS) is a progressive untreatable neurodegenerative disorder, leading to the death of the cortical and spinal motoneurons (MNs). Bone marrow-derived mesenchymal stem/stromal cells (BM-MSCs) may represent a new approach to slowing down the progression of ALS by providing neurotrophic support to host MNs and by having an anti-inflammatory effect. We have designed a prospective, nonrandomized, open-label clinical trial (phase I/IIa, EudraCT No. 2011-000362-35) to assess the safety and efficacy of autologous multipotent BM-MSCs in ALS treatment. Autologous BM-MSCs were isolated and expanded under GMP conditions. Patients received 15
± 4.5 ´ 106 of BM-MSCs via lumbar puncture into the cerebrospinal fluid. Patients were monitored for 6 months before treatment and then for an 18-month follow-up period. Potential adverse reactions were assessed, and the clinical outcome was evaluated by the ALS functional rating scale (ALSFRS), forced vital capacity (FVC), and weakness scales (WSs) to assess muscle strength on the lower and upper extremities. In total, 26 patients were enrolled in the study and were assessed for safety; 23 patients were suitable for efficacy evaluation. After intrathecal BM-MSC application, about 30% of the patients experienced a mild to moderate headache, resembling the headaches after a standard lumbar puncture. No suspected serious adverse reactions (SUSAR) were observed. We found a reduction in ALSFRS decline at 3 months after application ( p < 0.02) that, in some cases, persisted for 6 months ( p < 0.05). In about 80% of the patients, FVC values remained stable or above 70% for a time period of 9 months. Values of WS were stable in 75% of patients at 3 months after application. Our results demonstrate that the intrathecal application of BM-MSCs in ALS patients is a safe procedure and that it can slow down progression of the disease.

Key words: Clinical trial; Cell-based therapy; Stem cells; Amyotrophic lateral sclerosis (ALS) patients; Intrathecal application

Received September 15, 2016; final acceptance January 9, 2017. Online prepub date: November 7, 2016.
Address correspondence to Professor Eva Syková, M.D., Ph.D., Dr.Sc., Institute of Experimental Medicine, Czech Academy of Sciences, Videnska 1083, 142 20 Prague 4, Czech Republic. Tel: (+420 2) 4106 2204; Fax: (+420 2) 4106 2782; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 659-667, 2017
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DOI: https://doi.org/10.3727/096368916X
693059
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Step Sequence Is a Critical Gait Parameter of Unilateral 6-OHDA Parkinson’s Rat Models

Heather A. Baldwin,*1 Pyry P. Koivula,*1 Julie C. Necarsulmer,* Keith W. Whitaker,*†2 and Brandon K. Harvey*2

*Optogenetics and Transgenic Technology Core, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Baltimore, MD, USA
†Human Research and Engineering Directorate, US Army Research Laboratory, Aberdeen Proving Ground, MD, USA

Parkinson’s disease is a progressive neurological disorder, marked by the loss of dopaminergic neurons in the nigrostriatal pathway that leads to abnormal gait, rigidity, slowness of movement, and tremor. The ability to recapitulate and measure the neurological sequelae in rodent models of Parkinson’s disease is important for studying and evaluating potential therapeutics. Individual variability in lesion severity and injury progression are key factors in the 6-hydroxydopamine model that require normalization when evaluating therapeutic effects. The gait parameters that were found to be affected by 6-hydroxydopamine lesioning of the nigrostriatal pathway in rats may be used to study novel transgenic models of Parkinson’s disease as well as to test novel therapeutics. Previously, studies have used a video-based system to analyze gait abnormalities in the 6-hydroxydopamine model of Parkinson’s disease, but these studies did not account for individual variability on reported gait parameters. By analyzing the ratio of parameters from the injured to uninjured sides and correcting for speed in related parameters, hindpaw step cycle parameters, hindpaw print area, and step sequence are significantly altered in different ways for each type of lesion, when compared to saline-injected controls. These findings enable new metrics for evaluating therapeutic efficacy of drug-, gene-, or cell-based therapies in rat models of Parkinson’s disease.

Key words: CatWalk; Parkinson’s disease (PD); Gait analysis; Step sequence; 6-Hydroxydopamine (6-OHDA)

Received August 4, 2016; final acceptance November 18, 2016. Online prepub date: September 26, 2016.
1These authors provided equal contribution to this work.
2These authors provided equal contribution to this work.
Address correspondence to Brandon K. Harvey, NIH-NIDA-IRP-OTTC, Suite 200, Room 06A729, 251 Bayview Boulevard, Baltimore, MD 21224, USA. Tel: (443) 740-2590; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 669-677, 2017
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DOI: https://doi.org/10.3727/096368917X
695227
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Brilliant Blue G, But Not Fenofibrate, Treatment Reverts Hemiparkinsonian Behavior and Restores Dopamine Levels in an Animal Model of Parkinson’s Disease

Eneas G. Ferrazoli,*1 Hellio D.N. de Souza,†1 Isis C. Nascimento,† Agatha Oliveira-Giacomelli,† Telma T. Schwindt,† Luiz R. Britto,‡ and Henning Ulrich†

*Departamento de NeurologiaNeurocirurgiaDisciplina de Neurologia/NeurocienciaUniversidade Federal de Sao Paulo, Sao Paulo, Brazil
Departamento de Bioquimica, Instituto de QuimicaUniversidade de Sao Paulo, Sao Paulo, Brazil
Departamento de FisiologiaBiofisica, Instituto de Ciencias BiomedicasUniversidade de Sao Paulo, Sao Paulo, Brazil

Parkinson’s disease (PD) is a neurodegenerative disorder, characterized by the loss of dopaminergic neurons in the substantia nigra and their projections to the striatum. Several processes have been described as potential inducers of the dopaminergic neuron death, such as inflammation, oxidative stress, and mitochondrial dysfunction. However, the death of dopaminergic neurons seems to be multifactorial, and its cause remains unclear. ATP-activating purinergic receptors influence various physiological functions in the CNS, including neurotransmission. Purinergic signaling is also involved in pathological scenarios, where ATP is extensively released and promotes sustained purinergic P2X7 receptor (P2X7R) activation and consequent induction of cell death. This effect occurs, among other factors, by oxidative stress and during the inflammatory response. On the other hand, peroxisome proliferator-activated receptor-
g coactivator 1a (PGC-1a) is involved in energy metabolism and mitochondrial biogenesis. Expression and activity upregulation of this protein has been related with reduction of oxidative stress and neuroprotection. Therefore, P2X7R and PGC-1a are potential targets in the treatment of PD. Here hemiparkinsonism was induced by unilateral stereotactic injection of 6-OHDA in a rat model. After 7 days, the establishment of PD was confirmed and followed by treatment with the P2X7R antagonist Brilliant Blue G (BBG) or PGC-1a agonist fenofibrate. BBG, but not fenofibrate, reverted hemiparkinsonian behavior accompanied by an increase in tyrosine hydroxylase immunoreactivity in the substantia nigra. Our results suggest that the P2X7R may be a therapeutic target in Parkinson’s disease.

Key words: Parkinson’s disease (PD); P2X7 receptor; Brilliant Blue G (BBG); PGC-1a; Fenofibrate

Received October 3, 2015; final acceptance February 28, 2017. Online prepub date: March 3, 2017.
1These authors provided equal contribution to this work.
Address correspondence to Professor Henning Ulrich, Departamento de Bioquimica, Instituto de QuimicaUniversidade de Sao Paulo, Av. Prof. Lineu Prestes 748, Sao Paulo, SP 05508-900, Brazil. Tel: +55-11-3091-9622; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 679-691, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
693707
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved


A Subpopulation of Dopaminergic Neurons Coexpresses Serotonin in Ventral Mesencephalic Cultures But Not After Intrastriatal Transplantation in a Rat Model of Parkinson’s Disease

Stefano Di Santo,* Stefanie Seiler,* Angélique D. Ducray,* Morten Meyer,†‡ and Hans Rudolf Widmer*

*Department of Neurosurgery, Neurocenter and Regenerative Neuroscience Cluster, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland
†Department of Neurobiology Research, Institute of Molecular Medicine, University of Southern Denmark, Odense C, Denmark
‡Department of Neurology, Zealand University Hospital, Roskilde, Denmark

Cell replacement therapy is a promising avenue into the investigation and treatment of Parkinson’s disease (PD), and in some cases, significant long-term motor improvements have been demonstrated. The main source of donor tissue is the human fetal ventral mesencephalon (FVM), which consists of a mixed neuronal population, and its heterogeneity likely contributes to the inconsistent outcome observed in clinical trials. Therefore, detailed knowledge about the neuronal subpopulations in the VM seems essential for successful cell transplantation. Interestingly, it has been reported that some tyrosine hydroxylase-positive (TH+) neurons in the VM of adult rats and in cultured midbrain-derived neuroblasts coexpress additional neurotransmitters. Thus, the present study investigated, by means of colocalization analyses, the possible expression of GABA or serotonin in TH+ neurons. For that purpose, both fetal rat and human dissociated, organotypic and neurosphere FVM cultures as well as an animal model of PD were investigated. In dissociated rat FVM cultures, approximately 30% of the TH+ neurons coexpressed serotonin, while no colocalization with GABA was observed. Interestingly, coexpression of TH and serotonin was found to be dependent on the time in culture, the plating density, and the exposure to neurotrophic factors, that is, higher cell densities and treatment with brain-derived neurotrophic factor resulted in a significantly reduced coexpression rate. Notably, even though approximately 30% of the dopaminergic neurons in the donor tissue coexpressed serotonin, no colocalization could be detected in grafts 1 month after intrastriatal transplantation into hemiparkinsonian rats. In conclusion, a significant and susceptible subpopulation of dopaminergic neurons in FVM tissues coexpresses serotonin. This might have potential implications for the future selection and handling of cells prior to transplantation in PD.

Key words: Human; Tyrosine hydroxylase (TH); 5-Hydroxytryptamine (5HT); Cell transplantation; Parkinsonian rats

Received September 14, 2016; final acceptance December 13, 2016. Online prepub date: November 7, 2016.
Address correspondence to Hans R. Widmer, Ph.D., Department of Neurosurgery, University of Bern, OT O E215, Inselspital, 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


Cell Transplantation, Vol. 26, pp. 693-702, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
694184
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved


Regulator of Cell Cycle (RGCC) Expression During the Progression of Alzheimer’s Disease

Scott E. Counts*†‡ and Elliott J. Mufson§¶

*Department of Translational Science and Molecular Medicine, Michigan State University, Grand Rapids, MI, USA
†Department of Family Medicine, Michigan State University, Grand Rapids, MI, USA
Hauenstein Neurosciences Center, Mercy Health Saint Mary’s Hospital, Grand Rapids, MI, USA
§Department of Neurobiology, Barrow Neurological Institute, Phoenix, AZ, USA
¶Department of Neurology, Barrow Neurological Institute, Phoenix, AZ, USA

Unscheduled cell cycle reentry of postmitotic neurons has been described in cases of mild cognitive impairment (MCI) and Alzheimer’s disease (AD) and may form a basis for selective neuronal vulnerability during disease progression. In this regard, the multifunctional protein regulator of cell cycle (RGCC) has been implicated in driving G1/S and G2/M phase transitions through its interactions with cdc/cyclin-dependent kinase 1 (cdk1) and is induced by p53, which mediates apoptosis in neurons. We tested whether RGCC levels were dysregulated in frontal cortex samples obtained postmortem from subjects who died with a clinical diagnosis of no cognitive impairment (NCI), MCI, or AD. RGCC mRNA and protein levels were upregulated by ~50%–60% in MCI and AD compared to NCI, and RGCC protein levels were associated with poorer antemortem global cognitive performance in the subjects examined. To test whether RGCC might regulate neuronal cell cycle reentry and apoptosis, we differentiated neuronotypic PC12 cultures with nerve growth factor (NGF) followed by NGF withdrawal to induce abortive cell cycle activation and cell death. Experimental reduction of RGCC levels increased cell survival and reduced levels of the cdk1 target cyclin B1. RGCC may be a candidate cell cycle target for neuroprotection during the onset of AD.

Key words: Mild cognitive impairment (MCI); Alzheimer’s disease (AD); Cell cycle; Regulator of cell cycle (RGCC); Nerve growth factor (NGF); Apoptosis

Received September 29, 2016; final acceptance January 9, 2017. Online prepub date: November 30, 2016.
Address correspondence to Scott E. Counts, Ph.D., College of Human Medicine, Michigan State University, 333 Bostwick Avenue NE, Grand Rapids, MI 49503, USA. Tel: 616-234-0997; Fax: 616-234-0990: E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it