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3 Acta Neurobiologiae Experimentalis

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Transplantation of neurospheres and organoids derived from human umbilical cord blood onto hippocampal organotypic slice culture

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Sarnowska A. , Jurga M. , Domanska-Janik K.

nr 4

Heterogeneity of the local tissue microenvironment influences differentiation of NG2 cells

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Sypecka J. , Sarnowska A. , Szablowska-Gadomska I. , Lukomska D. , Domanska-Janik K.

nr 1

The NG2 cells are the oligodendrocyte precursors that terminally differentiated are capable for myelination of central nervous system (CNS). They exhibit many features of neural stem cells and constitute the abundant population of dividing progenitors in the young and adult brain. A question arises if their commitment could be modulated by local tissue-specific or neuropathological signals. The aim of our study therefore was to evaluate the effect of distinct microenvironments (provided by either the spinal cord or the hippocampal slices) on the differentiation of neonatal NG2 cells. Subsequently, hippocampal slice culture subjected to an ischemic injury (the glucose-oxygen deprivation, OGD) was used in order to evaluate the cell development in microenvironment conditioned by traumatized tissue. Methods. Both the hippocampal and spinal cord slice cultures were established from the same 7-day old rats. The model of an indirect contact (i.e. exclusively by the culture media) in co-culture system was chosen to eliminate the influence of cell-cell contact. The NG2 cells were obtained from 10-day old mixed primary culture of neonatal rat hemispheres. After 7 days in co-culture, the cells were either stained with neural markers or collected for the RNA isolation and real-time PCR. Results. The medium conditioned by hippocampal slices effectively promoted neurogenesis: ~30 % of NG2 cells differentiated into TUJ 1-positive neurons. The remaining fraction mostly formed premyelinating and mature oligodendrocytes. The exposition of hippocampal slices to the OGD injury abolished the effect of pro-neuronal induction in co-cultures. In media conditioned by spinal cord slices, neurogenesis was less pronounced (20% neurons) and the oligodendrocyte differentiation was significantly slowed-down. Conclusions. The NG2 cells were shown to have intrinsic potency for neurogenesis. Heterogeneity of local microenvironment might modify the fate of endogenous or transplanted NG2 cells what should be taken into consideration in potential neurorepair strategies. Supported by grant 0345/B/P01/2010/38.

MRI-monitored cord blood-derived cell transplantation to the ventricular system of child with global cerebral ischemia-a case report

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Janowski M. , Kmiec T. , Jurkiewicz E. , Kropiwnicki T. , Habich A. , Kotulska K. , Jelonek E. , Sarnowska A. , Litwin M. , Boruczkowski D. , Lukomska B. , Walecki J. , Roszkowski M. , Jozwiak S. , Domanska-Janik K.

nr 1

Introduction: Neurological disorders are the most common cause of serious disability and have a major impact on financial healthrelated burden to society. Most of them are definitely associated with cell death: sudden or chronic. Conventional treatment methods yield disappointing results. Thus the discoveries in stem cell biology have fueled the interest in cell-based therapeutical approach. Based on experimental data cord blood has been proposed as a novel, autologous cell source for pediatric population. Non-invasive monitoring of cell fate following transplantation has been recently recommended as a basis for rational stem cell therapy. Subject: One year old child experienced devastating, cardiac arrest-induced cerebral ischemia. Despite a broad rehabilitation program diagnose of vegetative state has been established three months later. After next three months of continued rehabilitation no noticeable improvement has also been found and the child has been included into study. The protocol has been approved by the ethical commission of The Children’s Memorial Health Institute in Warsaw, Poland. Then the child’s own cord blood cells have been neurally-converted over 10 days in culture within GMP facility. Prior to transplantation cells were labeled with iron oxide (SPIO) for MR imaging. For scaling sensitivity of MR signal different concentrations of SPIO-labeled cells were scanned in the phantom. Then patient received monthly 3 subsequent cell infusions (1.2 x 107 cells each) to lateral ventricles. The follow up continued up to 6 months and included both clinical assessment and MR examinations. Results: High efficiency of neural cell conversion and SPIO labeling as well as no cytotoxicity were observed. The employed method of cell transplantation was found to be efficient to deliver cells to CNS as confirmed by MR imaging. Gradual decrease of SPIO signal intensity was observed over the period of follow up. No adverse events or abnormal reaction to cell implantation was detected. The follow up revealed mild functional improvement - decreased nystagmus, spasticity and the number of epileptic seizures. Moreover, the features of the child contact with parents has appeared, thus vegetative state can not be diagnosed any more. Conclusions: This report indicates that transplantation of autologous, neurally-committed cord blood-derived cells to the ventricular system of child is safe, feasible and able to result with mild functional improvement. Additionally cell-related MRI signal can be monitored for more than 4 months in transplanted brain hemisphere. Supported by MSHE grants no 0141/B/P01/2008/35 and 0142/B/ P01/2008/35.