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1

Developmental response of human umbilical cord bIood-derived neural-like cells to selected neurotoxins

100%
Buzanska L., Sypecka J., Coecke S., Domanska-Janik K.
Acta Neurobiologiae Experimentalis
|
2006
|
tom 66
|
nr 4
2

Controlling of HUCB-NSC culture by micropatterned biofunctionized surfaces

76%
Zychowicz M., Buzanska L., Ruiz A., Coecke S., Rossi F., Domanska-Janik K.
Acta Neurobiologiae Experimentalis
|
2009
|
tom 69
|
nr 1
Stem cells have potential to maintain in organism by self-renewal division and ability to multilineage differentiation. Cell-cell contacts, paracrine signals and extracellular matrix proteins occurring in the neurogenic niches are main determinants of neural stem cells fate and dynamic of their differentiation. Aim of this study is to investigate whether could nano/micro-patterned, biofunctionalized surface guides human cord blood derived neural stem cell to growth and differentiation. Methods: To investigate the infl uence of the cell plating density we used the microcontact printed patterns of adhesive substrate (poly-L-lysine) on cell-repellent poly-ethylene glycol (PEG) substrate. Two different geometries of the patterning have been applied: to test proliferative response the cells were seeded in different densities on the pattern with separated pitches, while to verify their ability to differentiation the culture medium was supplemented with cAMP and cells seeded on the surface patterned with interconnecting lines. After 2, 4 and 7 days the cells were fi xed and immunostained for Ki67 (proliferation marker) and beta III tubulin/ S100β markers for neuronal/astrocytic lineage. Results: The low cell density of HUCB-NSC (104 cell/cm2 ) and the presence of neuromorphogenes (cAMP) supports neural stem cell differentiation, while enhanced initial cell density promotes the growth rate (increase of the cell number falling on biofunctionalized unit per 100 μm2 surface). Conclusions: Micropatterned platforms with biofunctionalized surface can be used for screening of the plethora of extracellular signals directing neural stem cell to growth and differentiation.
3

Expression of the membrane complement regulatory proteins [CD55 and CD59] in human thymus

76%
Sladowski D., Wasiutynski A., Wilczynski G., Grabska-Liberek I., Coecke S., Kinsner A., Kochanowska I.
Folia Histochemica et Cytobiologica
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2006
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tom 44
|
nr 4
4

Human cord blood cells in neurotoxicology: Advantage of emerging technologies

64%
Buzanska L., Zychowicz M., Ruiz A., Ceriotti L., Sobanski T., Rossi F., Calpo P., Domanska-Janik K., Coecke S.
Acta Neurobiologiae Experimentalis
|
2009
|
tom 69
|
nr 1
Stem cell technology provides a new tool for better understanding the mechanisms involved in compound-induced adverse reactions of the organism, which particularly applies to the fi eld of developmental neurotoxicity. Human Umbilical Cord Blood Neural Stem Cell (HUCB-NSC) line is a model system where key neurodevelopmental processes were investigated by conventional and emerging techniques. The advantage of the HUCB-NSC line is that cells are of human origin, non-transformed and can be cultured/harvested at different developmental stages. In this report emerging nano/micro-technologies were used to create biofunctional micropatterns and multi electrode array chips for the detection of cell behaviour and vulnerability to toxic compounds. Micropatterned surfaces were produced by a spatial arrangement of different functional domains. This included a nano/micro-fabrication technique like contact printing in order to create a pattern of separated or interconnected polypeptide spots directing cell growth and differentiation. Another approach was to create protein microarrays by piezoelectric (non contact) deposition of extracellular matrix proteins. Such a method allows defi ned active areas to be produced on the same platform and enables unambiguous access to cell behavior on different protein types and concentrations. HUCB-NSC were shown to adhere and differentiate on microarray platforms in a protein type, concentration and cell density dependent manner. Sensor and omics techniques applied to HUCB-NSC included measurements of electrical activity using multielectrode array chips and metabolite profi ling by mass spectrometry. Spontaneous electrical fi eld potentials and the protein composition of tested cells were sensitive to neurotoxic treatments in a developmental stage specifi c manner.
5

Patterned growth and differentiation of human cord blood-derived neural stem cells on bio-functionalized surfaces

64%
Buzanska L., Ruiz A., Zychowicz M., Rauscher H., Ceriotti L., Rossi F., Colpo P., Domanska-Janik K., Coecke S.
Acta Neurobiologiae Experimentalis
|
2009
|
tom 69
|
nr 1
Bio-functionalized surfaces were prepared to study the adherence and differentiation capacity of neural stem cells derived from human umbilical cord blood (HUCB-NSC). Cell growth platforms containing arranged arrays of adhesive molecules were created by microcontact printing on a biologically inert surface. Biomolecules used to prepare microarray platforms included the extracellular matrix protein fibronectin and the polyaminoacid poly-L-lysine. HUCB-NSC plated on microplatforms at various serum conditions showed serum and molecule type dependent capacity for adhesion and differentiation. Poly-L-lysine allowed the maintenance of stem-like non differentiated cells attached to the surface, whereas fibronectin promoted spreading and neural commitment. Serum deprivation did not influence the attachment of HUCB-NSC to fibronectin, but significantly enhanced the attachment to poly-L-lysine and promoted dBcAMP induced neuronal differentiation. A bio-pattern of squares with interconnecting lines was used to guide neuronal differentiation by directing cell protrusion outgrowth. Tailoring the geometry of the bio-pattern enabled directing and monitoring of the neural stem cells development in the large scale multiparameter biotests.
6

HUCB-NSCS growth control by biofunctionalized surfaces

52%
Zychowicz M., Ruiz A., Cerioti L., Lisboa P., Mehn D., Coecke S., Rausher H., Colpo P., Rossi F., Domanska-Janik K., Buzanska L.
Acta Neurobiologiae Experimentalis
|
2009
|
tom 69
|
nr 3
Cell growth platforms with biofunctionalized surfaces were fabricated to control and direct HUCB-NSC fate decisions. Two different nano/micro techniques: microcontact printing and piezoelectric non-contact spotting were used to allocate biomolecules (poly-Llysine and fi bronectin) on cell-repellent, non adhesive substrate. Such methods allow controlling the spatial distribution and content of the biomolecules on the microarray and governing cell adhesion in unspecifi c (electrostatic) or specifi c (receptor-mediated) manner. Patterning of biomolecules in different conditions on the single growth platforms enables to infl uence and compare stem cells developmental processes (proliferation /differentiation) at variable environments. To refl ect/mimic stem cell niche we applied functional domains containing ECM protein spotted together with the small signaling molecules (notch, wnt, shh). Such approach enable directing of neural stem cell developmental program by inducing intracellular molecular pathways leading to either self renewing- or differentiating- (neuronal or astrocytic) state. We characterized active biofunctionalized domains on fabricated microarrays by applying two methods of surface analysis: ellipsometry measurement and surface plasmon resonance system. This type of bioengineered cell growth platforms can be used for screening the mechanisms governing neural stem cell fate decisions and adverse reactions upon environmental stimuli. Grant No 0141/B/ P01/2008/35.
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