Publications

Hydrogel-based scaffolds to support intrathecal stem cell transplantation as a gateway to the spinal cord: clinical needs, biomaterials, and imaging technologies.

NPJ Regen Med. 2018 Apr 4;3:8. doi: 10.1038/s41536-018-0046-3. eCollection 2018.

Oliveira JM1,2,3, Carvalho L1,2, Silva-Correia J1,2, Vieira S1,2, Majchrzak M4, Lukomska B4, Stanaszek L4, Strymecka P4, Malysz-Cymborska I5, Golubczyk D5, Kalkowski L5, Reis RL1,2,3, Janowski M4,6,7, Walczak P5,6,7.

1 3B´s Research Group - Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence, Tissue Engineering and Regenerative Medicine, AvePark, Zona Industrial da Gandra, 4805-017 Barco, Guimarães Portugal.

2 2ICVS/3B's - PT Government Associate Laboratory, Braga, Portugal.

3 3The Discoveries Centre for Regenerative and Precision Medicine, Headquarters at University of Minho, Avepark, 4805-017 Barco, Guimarães Portugal.

4 4NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.

5 5Department of Neurology and Neurosurgery, School of Medicine, Collegium Medicum, University of Warmia and Mazury, Olsztyn, Poland.

6 6Russel H, Morgan Department of Radiology and Radiological Science, Johns Hopkins University, Baltimore, MD USA.

7 7Vascular Biology Program, Institute for Cell Engineering, Johns Hopkins University, Baltimore, MD USA.

Hydrogel-based scaffolds to support intrathecal stem cell transplantation as a gateway to the spinal cord: clinical needs, biomaterials, and imaging technologies.

ABSTRACT

The prospects for cell replacement in spinal cord diseases are impeded by inefficient stem cell delivery. The deep location of the spinal cord and complex surgical access, as well as densely packed vital structures, question the feasibility of the widespread use of multiple spinal cord punctures to inject stem cells. Disorders characterized by disseminated pathology are particularly appealing for the distribution of cells globally throughout the spinal cord in a minimally invasive fashion. The intrathecal space, with access to a relatively large surface area along the spinal cord, is an attractive route for global stem cell delivery, and, indeed, is highly promising, but the success of this approach relies on the ability of cells (1) to survive in the cerebrospinal fluid (CSF), (2) to adhere to the spinal cord surface, and (3) to migrate, ultimately, into the parenchyma. Intrathecal infusion of cell suspension, however, has been insufficient and we postulate that embedding transplanted cells within hydrogel scaffolds will facilitate reaching these goals. In this review, we focus on practical considerations that render the intrathecal approach clinically viable, and then discuss the characteristics of various biomaterials that are suitable to serve as scaffolds. We also propose strategies to modulate the local microenvironment with nanoparticle carriers to improve the functionality of cellular grafts. Finally, we provide an overview of imaging modalities for in vivo monitoring and characterization of biomaterials and stem cells. This comprehensive review should serve as a guide for those planning preclinical and clinical studies on intrathecal stem cell transplantation.

The Differentiation of Rat Oligodendroglial Cells Is Highly Influenced by the Oxygen Tension: In Vitro Model Mimicking Physiologically Normoxic Conditions.

Int J Mol Sci. 2018 Jan 24;19(2). pii: E331. doi: 10.3390/ijms19020331.

Janowska J1, Ziemka-Nalecz M2, Sypecka J3.

1 NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences, 5 Pawinskiego Street, 02-106 Warsaw, Poland. This email address is being protected from spambots. You need JavaScript enabled to view it..

2 NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences, 5 Pawinskiego Street, 02-106 Warsaw, Poland. This email address is being protected from spambots. You need JavaScript enabled to view it..

3 NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences, 5 Pawinskiego Street, 02-106 Warsaw, Poland. This email address is being protected from spambots. You need JavaScript enabled to view it..

The Differentiation of Rat Oligodendroglial Cells Is Highly Influenced by the Oxygen Tension: In Vitro Model Mimicking Physiologically Normoxic Conditions.

ABSTRACT

Oligodendrocyte progenitor cells (OPCs) constitute one of the main populations of dividing cells in the central nervous system (CNS). Physiologically, OPCs give rise to mature, myelinating oligodendrocytes and confer trophic support to their neighboring cells within the nervous tissue. OPCs are known to be extremely sensitive to the influence of exogenous clues which might affect their crucial biological processes, like survival, proliferation, differentiation, and the ability to generate a myelin membrane. Alterations in their differentiation influencing their final potential for myelinogenesis are usually the leading cause of CNS dys- and demyelination, contributing to the development of leukodystrophic disorders. The evaluation of the mechanisms that cause oligodendrocytes to malfunction requires detailed studies based on designed in vitro models. Since OPCs readily respond to changes in local homeostasis, it is crucial to establish restricted culture conditions to eliminate the potential stimuli that might influence oligodendrocyte biology. Additionally, the in vitro settings should mimic the physiological conditions to enable the obtained results to be translated to future preclinical studies. Therefore, the aim of our study was to investigate OPC differentiation in physiological normoxia (5% O₂) and a restricted in vitro microenvironment. To evaluate the impact of the combined microenvironmental clues derived from other components of the nervous tissue, which are also influenced by the local oxygen concentration, the process of generating OPCs was additionally analyzed in organotypic hippocampal slices. The obtained results show that OPC differentiation, although significantly slowed down, proceeded correctly through its typical stages in the physiologically relevant conditions created in vitro. The established settings were also conducive to efficient cell proliferation, exerting also a neuroprotective effect by promoting the proliferation of neurons. In conclusion, the performed studies show how oxygen tension influences OPC proliferation, differentiation, and their ability to express myelin components, and should be taken into consideration while planning preclinical studies, e.g., to examine neurotoxic compounds or to test neuroprotective strategies.

KEYWORDS:

cell proliferation; culture density; hippocampal organotypic slices; myelin protein amounts; myelinogenesis; oligodendrocyte maturation myelin; oligodendrocyte progenitor cells; physiological normoxia; serum-free culture

Impact of neonatal hypoxia-ischaemia on oligodendrocyte survival, maturation and myelinating potential.

J Cell Mol Med. 2018 Jan;22(1):207-222. doi: 10.1111/jcmm.13309. Epub 2017 Aug 7.

Ziemka-Nalecz M1, Janowska J1, Strojek L1, Jaworska J1, Zalewska T1, Frontczak-Baniewicz M2, Sypecka J1.

Author information

1 NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.

2 Electron Microscopy Platform, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.

 

Impact of neonatal hypoxia-ischaemia on oligodendrocyte survival, maturation and myelinating potential.

ABSTRACT

Hypoxic-ischaemic episodes experienced at the perinatal period commonly lead to a development of neurological disabilities and cognitive impairments in neonates or later in childhood. Clinical symptoms often are associated with the observed alterations in white matter in the brains of diseased children, suggesting contribution of triggered oligodendrocyte/myelin pathology to the resulting disorders. To date, the processes initiated by perinatal asphyxia remain unclear, hampering the ability to develop preventions. To address the issue, the effects of temporal hypoxia-ischaemia on survival, proliferation and the myelinating potential of oligodendrocytes were evaluated ex vivo using cultures of hippocampal organotypic slices and in vivo in rat model of perinatal asphyxia. The potential engagement of gelatinases in oligodendrocyte maturation was assessed as well. The results pointed to a significant decrease in the number of oligodendrocyte progenitor cells (OPCs), which is compensated for to a certain extent by the increased rate of OPC proliferation. Oligodendrocyte maturation seemed however to be significantly altered. An ultrastructural examination of selected brain regions performed several weeks after the insult showed however that the process of developing central nervous system myelination proceeds efficiently resulting in enwrapping the majority of axons in compact myelin. The increased angiogenesis in response to neonatal hypoxic-ischaemic insult was also noticed. In conclusion, the study shows that hypoxic-ischaemic episodes experienced during the most active period of nervous system development might be efficiently compensated for by the oligodendroglial cell response triggered by the insult. The main obstacle seems to be the inflammatory process modulating the local microenvironment.

KEYWORDS:

electron microscopy; gelatinases; hippocampal organotypic slices; myelin structure; myelinogenesis; neonatal hypoxia-ischaemia; oligodendrocyte progenitor cells; oxygen and glycose deprivation; perinatal asphyxia

Imaging of extracellular vesicles derived from human bone marrow mesenchymal stem cells using fluorescent and magnetic labels

Int J Nanomedicine. 2018 Mar 19;13:1653-1664. doi: 10.2147/IJN.S159404. eCollection 2018.

Dabrowska S1, Del Fattore A2, Karnas E3,4, Frontczak-Baniewicz M5, Kozlowska H6, Muraca M7, Janowski M1,8, Lukomska B1.

1 NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.

2 Multifactorial Disease and Complex Phenotype Research Area, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy.

3 Department of Cell Biology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.

4 Malopolska Centre of Biotechnology, Krakow, Poland.

5 Electron Microscopy Platform, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.

6 Laboratory of Advanced Microscopy Techniques, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.

7 Department of Women's and Children's Health, University of Padua, Padua, Italy.

8 Russel H. Morgan Department of Radiology and Radiological Science, Division of MR Research, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.

Imaging of extracellular vesicles derived from human bone marrow mesenchymal stem cells using fluorescent and magnetic labels.

 

ABSTRACT

Mesenchymal stem cells have been shown therapeutic in various neurological disorders. Recent studies support the notion that the predominant mechanism by which MSCs act is through the release of extracellular vesicles (EVs). EVs seem to have similar therapeutic activity as their cellular counterparts and may represent an interesting alternative standalone therapy for various diseases. The aim of the study was to optimize the method of EV imaging to better understand therapeutic effects mediated by EVs.

 

KEYWORDS:

MRI; cell tracking; extracellular vesicles; fluorescent dye; iron oxide; mesenchymal stem cells

Glycogen metabolism in brain and neurons - astrocytes metabolic cooperation can be altered by pre- and neonatal lead (Pb) exposure.

Toxicology. 2017 Sep 12;390:146-158. doi: 10.1016/j.tox.2017.09.007. 

Baranowska-Bosiacka I1, Falkowska A2, Gutowska I3, Gąssowska M4, Kolasa-Wołosiuk A5, Tarnowski M6, Chibowska K2, Goschorska M2, Lubkowska A7, Chlubek D2.
Author information
Glycogen metabolism in brain and neurons - astrocytes metabolic cooperation can be altered by pre- and neonatal lead (Pb) exposure.
1 Department of Biochemistry and Medical Chemistry, Powstańców Wlkp. 72 St., 70-111 Szczecin, Pomeranian Medical University in Szczecin, Poland. Electronic address: This email address is being protected from spambots. You need JavaScript enabled to view it..
2 Department of Biochemistry and Medical Chemistry, Powstańców Wlkp. 72 St., 70-111 Szczecin, Pomeranian Medical University in Szczecin, Poland.
3 Department of Biochemistry and Human Nutrition, Broniewskiego 24 St., 71-460 Szczecin, Pomeranian Medical University in Szczecin, Poland.
4 Department of Cellular Signalling, Mossakowski Medical Research Centre, Polish Academy of Sciences, Pawińskiego 5, 02-106 Warsaw, Poland.
5 Department of Histology and Embriology, Powstańców Wlkp. 72 St., 70-111 Szczecin, Pomeranian Medical University in Szczecin, Poland.
6 Department of Physiology, Powstańców Wlkp. 72 St., 70-111 Szczecin, Pomeranian Medical University in Szczecin, Poland.
7 Department of Functional Diagnostics and Physical Medicine, Pomeranian Medical University in Szczecin, Żołnierska 48 Str, 71-210 Szczecin, Poland.

ABSTRACT

Lead (Pb) is an environmental neurotoxin which particularly affects the developing brain but the molecular mechanism of its neurotoxicity still needs clarification. The aim of this paper was to examine whether pre- and neonatal exposure to Pb (concentration of Pb in rat offspring blood below the "threshold level") may affect the brain's energy metabolism in neurons and astrocytes via the amount of available glycogen. We investigated the glycogen concentration in the brain, as well as the expression of the key enzymes involved in glycogen metabolism in brain: glycogen synthase 1 (Gys1), glycogen phosphorylase (PYGM, an isoform active in astrocytes; and PYGB, an isoform active in neurons) and phosphorylase kinase β (PHKB). Moreover, the expression of connexin 43 (Cx43) was evaluated to analyze whether Pb poisoning during the early phase of life may affect the neuron-astrocytes' metabolic cooperation. This work shows for the first time that exposure to Pb in early life can impair brain energy metabolism by reducing the amount of glycogen and decreasing the rate of its metabolism. This reduction in brain glycogen level was accompanied by a decrease in Gys1 expression. We noted a reduction in the immunoreactivity and the gene expression of both PYGB and PYGM isoform, as well as an increase in the expression of PHKB in Pb-treated rats. Moreover, exposure to Pb induced decrease in connexin 43 immunoexpression in all the brain structures analyzed, both in astrocytes as well as in neurons. Our data suggests that exposure to Pb in the pre- and neonatal periods results in a decrease in the level of brain glycogen and a reduction in the rate of its metabolism, thereby reducing glucose availability, which as a further consequence may lead to the impairment of brain energy metabolism and the metabolic cooperation between neurons and astrocytes.

KEYWORDS:

Brain glycogen metabolism; Glycogen phosphorylase brain isoform (PYGB); Glycogen phosphorylase kinase (PHKB); Glycogen phosphorylase muscle isoform (PYGM); Glycogen synthase (Gys1); Lead (Pb) neurotoxicity

Phenotypic, Functional, and Safety Control at Preimplantation Phase of MSC-Based Therapy.

Stem Cells Int. 2016;2016:2514917. doi: 10.1155/2016/2514917. Epub 2016 Aug 29.

Lech W1, Figiel-Dabrowska A1, Sarnowska A1, Drela K1, Obtulowicz P1, Noszczyk BH2, Buzanska L1, Domanska-Janik K1.

Phenotypic, Functional, and Safety Control at Preimplantation Phase of MSC-Based Therapy.

Author information

1 Mossakowski Medical Research Centre, Polish Academy of Sciences, 5 Pawinskiego Street, 02-106 Warsaw, Poland.

2 Department of Plastic Surgery at the Medical Center for Postgraduate Education, Warsaw School of Medicine, 01-828 Warsaw, Poland.

 

ABSTRACT

Mesenchymal stem cells (MSC) exhibit enormous heterogeneity which can modify their regenerative properties and therefore influence therapeutic effectiveness as well as safety of these cells transplantation. In addition the high phenotypic plasticity of MSC population makes it enormously sensitive to any changes in environmental properties including fluctuation in oxygen concentration. We have shown here that lowering oxygen level far below air atmosphere has a beneficial impact on various parameters characteristic for umbilical cord Wharton Jelly- (WJ-) MSC and adipose tissue- (AD-) derived MSC cultures. This includes their cellular composition, rate of proliferation, and maintenance of stemness properties together with commitment to cell differentiation toward mesodermal and neural lineages. In addition, the culture genomic stability increased significantly during long-term cell passaging and eventually protected cells against spontaneous transformation. Also by comparing of two routinely used methods of MSCs isolation (mechanical versus enzymatic) we have found substantial divergence arising between cell culture properties increasing along the time of cultivation in vitro. Thus, in this paper we highlight the urgent necessity to develop the more sensitive and selective methods for prediction and control cells fate and functioning during the time of growth in vitro.

Sensitivity of hiPSC-derived neural stem cells (NSC) to Pyrroloquinoline quinone depends on their developmental stage.

Toxicol In Vitro. 2017 May 31. pii: S0887-2333(17)30133-9. doi: 10.1016/j.tiv.2017.05.017. [Epub ahead of print]

Sensitivity of hiPSC-derived neural stem cells (NSC) to Pyrroloquinoline quinone depends on their developmental stage.

Augustyniak J1, Lenart J2, Zychowicz M1, Lipka G1, Gaj P3, Kolanowska M4, Stepien PP5, Buzanska L6.

Author information

1 Stem Cell Bioengineering Unit, Mossakowski Medical Research Centre, Polish Academy of Sciences, Poland.

2 Department of Neurochemistry, Mossakowski Medical Research Centre, Polish Academy of Sciences, Poland.

3 Laboratory of Human Cancer Genetics, Centre of New Technologies, CENT, University of Warsaw, Warsaw, Poland.

4 Laboratory of Human Cancer Genetics, Centre of New Technologies, CENT, University of Warsaw, Warsaw, Poland; Genomic Medicine, Medical University of Warsaw, Warsaw, Poland.

5 Department of Genetics, Faculty of Biology, University of Warsaw, Poland; Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw; Centre for New Technologies, University of Warsaw, Poland.

6 Stem Cell Bioengineering Unit, Mossakowski Medical Research Centre, Polish Academy of Sciences, Poland. Electronic address: This email address is being protected from spambots. You need JavaScript enabled to view it..

ABSTRACT

Pyrroloquinoline quinone (PQQ) is a factor influencing on the mitochondrial biogenesis. In this study the PQQ effect on viability, total cell number, antioxidant capacity, mitochondrial biogenesis and differentiation potential was investigated in human induced Pluripotent Stem Cells (iPSC) - derived: neural stem cells (NSC), early neural progenitors (eNP) and neural progenitors (NP). Here we demonstrated that sensitivity to PQQ is dependent upon its dose and neural stage of development. Induction of the mitochondrial biogenesis by PQQ at three stages of neural differentiation was evaluated at mtDNA, mRNA and protein level. Changes in NRF1, TFAM and PPARGC1A gene expression were observed at all developmental stages, but only at eNP were correlated with the statistically significant increase in the mtDNA copy numbers and enhancement of SDHA, COX-1 protein level. Thus, the "developmental window" of eNP for PQQ-evoked mitochondrial biogenesis is proposed. This effect was independent of high antioxidant capacity of PQQ, which was confirmed in all tested cell populations, regardless of the stage of hiPSC neural differentiation. Furthermore, a strong induction of GFAP, with down regulation of MAP2 gene expression upon PQQ treatment was observed. This indicates a possibility of shifting the balance of cell differentiation in the favor of astroglia, but more research is needed at this point.

KEYWORDS:

Developmental neurotoxicity; Mitochondrial biogenesis; Neural progenitors; Neural stem cells; PQQ; hiPSC

Epigenetic Modulation of Stem Cells in Neurodevelopment: The Role of Methylation and Acetylation.

Front Cell Neurosci. 2017 Feb 7;11:23. doi: 10.3389/fncel.2017.00023. eCollection 2017.

Podobinska M1, Szablowska-Gadomska I2, Augustyniak J1, Sandvig I3, Sandvig A3, Buzanska L1.

Epigenetic Modulation of Stem Cells in Neurodevelopment: The Role of Methylation and Acetylation.

Author information

1 Stem Cell Bioengineering Unit, Mossakowski Medical Research Centre, Polish Academy of Sciences Warsaw, Poland.

2 Pharmacology Department, Institute of Mother and Child Warsaw, Poland.

3 Department of Neuromedicine and Movement Science, Faculty of Medicine and Health Sciences, Norwegian University of Science and Technology (NTNU) Trondheim, Norway.

ABSTRACT

The coordinated development of the nervous system requires fidelity in the expression of specific genes determining the different neural cell phenotypes. Stem cell fate decisions during neurodevelopment are strictly correlated with their epigenetic status. The epigenetic regulatory processes, such as DNA methylation and histone modifications discussed in this review article, may impact both neural stem cell (NSC) self-renewal and differentiation and thus play an important role in neurodevelopment. At the same time, stem cell decisions regarding fate commitment and differentiation are highly dependent on the temporospatial expression of specific genes contingent on the developmental stage of the nervous system. An interplay between the above, as well as basic cell processes, such as transcription regulation, DNA replication, cell cycle regulation and DNA repair therefore determine the accuracy and function of neuronal connections. This may significantly impact embryonic health and development as well as cognitive processes such as neuroplasticity and memory formation later in the adult.

KEYWORDS:

acetylation; epigenetics; methylation; neurodevelopment; neuroplasticity; stem cells

Concise Review: MSC Adhesion Cascade-Insights into Homing and Transendothelial Migration.

Stem Cells. 2017 Jun;35(6):1446-1460. doi: 10.1002/stem.2614. Epub 2017 Apr 3.

Nitzsche F1,2, Müller C1, Lukomska B3, Jolkkonen J4, Deten A5, Boltze J1,5,6.

Concise Review: MSC Adhesion Cascade-Insights into Homing and Transendothelial Migration.

Author information:

1 Department of Ischemia Research, Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany.

2 Department of Radiology, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.

3 NeuroRepair Department, Mossakowski Medical Research Centre, Warsaw, Poland.

4 Department of Neurology, Institute of Clinical Medicine, University of Eastern, Kuopio, Finland.

5 Translational Centre for Regenerative Medicine, Leipzig University, Leipzig, Germany.

6 Department of Translational Medicine and Cell Technology, Fraunhofer Research Institution for Marine Biotechnology and Institute for Medical and Marine Biotechnology, University of Lübeck, Lübeck, Germany.

 

ABSTRACT

Mesenchymal stem cells (MSCs) are promising candidates for adult cell therapies in regenerative medicine. To fully exert their potential, efficient homing and migration toward lesion sites play an important role. Local transplantation deposits MSC in spatial proximity to the lesion, but often requires invasive procedures. Systemic administration routes are favored, but require the targeted extravasation of the circulating MSC at the site of injury. Transplanted MSC can indeed leave the blood flow and transmigrate through the endothelial barrier, and reach the lesion site. However, the underlying processes are not completely dissolved yet. Recent in vitro and in vivo research identified some key molecules scattered light on the extravasation mechanism. This review provides a detailed overview over the current knowledge of MSC transendothelial migration. We use the leukocyte extravasation process as a role model to build a comprehensive concept of MSC egress mechanisms from the blood stream and identified relevant similarities as well as important differences between the extravasation mechanisms. Stem Cells 2017;35:1446-1460.

KEYWORDS:

Cell adhesion molecules; Cell migration; Chemokine receptors; Integrins; Leukocytes; Mesenchymal stem cells; Stem/progenitor cel.