Preferencje help
Polish version English version Język
Widoczny [Schowaj] Abstrakt
Liczba wyników

Znaleziono wyników: 17

Liczba wyników na stronie
Pierwsza strona wyników Pięć stron wyników wstecz Poprzednia strona wyników Strona / 1 Następna strona wyników Pięć stron wyników wprzód Ostatnia strona wyników

Wyniki wyszukiwania

help Sortuj według:

help Ogranicz wyniki do:
Pierwsza strona wyników Pięć stron wyników wstecz Poprzednia strona wyników Strona / 1 Następna strona wyników Pięć stron wyników wprzód Ostatnia strona wyników
1

Bioactive domains controlling neural stem cells fate decisions

100%
Zychowicz M., Buzanska L.
Acta Neurobiologiae Experimentalis
|
2011
|
tom 71
|
nr S
Studying in vitro mutual interactions of the stem cells with the components of their microenvironment mimicking in vivo conditions is crucial for the further tissue engineering and regenerative medicine applications. Bioactive domains obtained by nano/micro emerging technologies, such as microcontact printing or piezoelectric spotting, were designed to reflect the stem cell niche composition and to influence their fate decisions. Different geometry of biofunctionalized surfaces, obtained by microcontact printing of poly-L-lysine or fibronectin, was created to study the adhesion, migration, proliferation as well as the differentiation of human cord blood-derived neural stem cells (HUCB-NSCs). The bioactive domains microspotted on the cell repellant surface contained extracellular matrix protein - fibronectin enriched with small signaling molecules, (CNTF, Jagged, Wnt, Shh, Dkk) were designed to activate particular molecular pathways leading to the maintenance of the self renewal of non-differentiated cells or to promote their differentiation into different neural lineages. Our results revealed how to control neural stem cell fate decisions by manipulating with the architecture and composition of the niche bioactive domains in vitro. Moreover, such domains can be used to investigate the stem cell response to cyto- or neurotoxins (MeHgCl), thus establishing good tool for screening the effect of diverse factors influencing neural stem cell development. Sponsored by Polish Ministry of Scientific Research and Higher Education grant No: 2211/B/P01/2010/38 and European Commission Joint Research Centre NanoBioscience Action
2

Biomimetic modulation of cord blood derived stem cells for neural fate control

100%
Buzanska L., Zychowicz M.
Acta Neurobiologiae Experimentalis
|
2011
|
tom 71
|
nr 1
Bioactive surface domains were applied to investigate cellular developmental processes of human cord blood- derived stem cells and to direct their fate into desired neural lineages. Such domains should represent microenvironmental cues resembling those found in vivo. For that purpose we have created miniaturized cell growth platforms with defined arrays of cell attractive biomaterials serving as functional domains. Emerging technologies applied included a nano/micro-fabrication technique like microcontact printing and piezoelectric (noncontact) microspotting of biomolecules on plasma deposited cell repellent surface. Human Umbilical Cord Blood Neural Stem Cell (HUCB-NSC) line was plated on biodomains at different concentrations and serum conditions. HUCB-NSCs were shown to adhere and differentiate on microarray platforms in a protein type, concentration and cell density dependent manner. Receptor-mediated interactions with extracellular proteins promote neuronal differentiation, while non-specific adhesion to polyaminoacid molecules allows maintaining of stem cells immobilized to the surface in non-differentiated stage. “Smart” functional domains were created by immobilizing to the surface small signaling molecules (e.g wnt, shh, notch or jagged) together with ECM proteins. Stimulation of selected intracellular pathways by signaling molecules resulted in differentiation of HUCB-NSC to either neuronal or astroglial lineage. Miniaturization of such bioengineered active domains combined with appropriate stem cell model may allow application of such stem cell growth platforms for the multiparameter bio-tests and can provide important, additional information on the sensitivity of certain neural stem cell molecular pathways to the selected neurotoxins. Since HUCB-NSC can be cultured and harvested at different developmental stages and was shown to be a good model for developmental toxicity testing, homogenous lineage related pluripotent population is required. For that purpose iPs cells from HUCB-NSC are produced. Sponsored by grant from Polish Ministry of Scientific Research and Higher Education No 0141/B/P01/2008/35, No N N302 597838. and European Commission, JRC.
3

Examination of the microscopic imaging of neural stem cells for counting a number of cells

81%
Korzrynska A., Dobrowolska E., Zychowicz M., Hoser P.
Acta Neurobiologiae Experimentalis
|
2009
|
tom 69
|
nr 1
The goal of the investigation is to examine cells imaging with the brightfi eld and confocal microscopic imaging using two techniques: with and without fl uorescence. The investigations are done as an introducing part of the long term goal to develop methods of monitoring a number of cells in culture in time. Several types of events in the monitored cellsí culture, e. g. cellís division and death, cellsí fusion and its changes in shape and/or position should be detected and quantifi ed. Because of immanent features of each microscopic technique some of events are easier to be observed in one of the microscopic techniques than in others. Other events are detectable in all microscopic imaging. To detect them, the area covered by the cells in the image should be selected in the fi rst step of the image processing. So the method of cellsí area detection, described by Korzynska and Iwanowski (2008) [In: Pietka, Kawa (Eds), Information Technique in Biomedicine, ASC 47, pp. 365ñ376], is applied to all tested types of the microscopic images of HUCB-NSC transfected by GFP (from Medical Research Center PAS) and the results of area detection are examined and compared. It was found, that the proposed method detects cellsí area more precisely in the fl uorescent images, than in the brightfi eld or confocal microscopy. Comparing results of cellsí area detection of the fl uorescent image of confocal microscopy with the integrated fl uorescent signal in epifl uorescent images of conventional optical microscopy the better result of detection cells in the image plane were observed for fl attened, large cells in confocal images while for rounded, convergent ones for the brightfi eld microscopic techniques.
4

Wpływ pastwiska na stan zarażenia bydła pasożytami przewodu poakrmowego

81%
Nowosad B., Skalska M., Fudalewicz-Niemczyk W., Zychowicz M.
Zeszyty Naukowe Akademii Rolniczej w Krakowie. Zootechnika
|
1996
|
tom 31
5

Sensitivity of neural stem cells derived from human umbilical cord blood (HUCB-NSC) to developmental neurotoxin - methylmercury chloride: Dependence on non-specific and receptor mediated interactions with biomolecules

81%
Dziedzicka D., Zychowicz M., Stepien P., Buzanska L.
Acta Neurobiologiae Experimentalis
|
2013
|
tom 73
|
nr 1
Human neural stem cells play an important role in in vitro developmental neurotoxicity testing. The purpose of this research was to investigate the sensitivity of neural stem cells derived from human umbilical cord blood (HUCB-NSC) to methylmercury chloride (MeHgCl), and its dependence on the type of interaction on cell membrane/biomolecule interface. MeHgCl is well known neurotoxin with documented adverse influence on human central nervous system (CNS) development. Cells were cultured in 96-well plates covered with different adhesive substrates or on Petri dishes microcontact-printed with biofunctional domains. The following biomaterials were used: poly-L-lysine, the synthetic compound, which allows to create electrostatic interactions with cells, or fibronectin and vitronectin, proteins of extracellular matrix, which create receptor mediated interactions between cells and the adhesive substrate. After the incubation with different concentrations of the neurotoxin, the cell viability, ability to proliferate, and to differentiate into neural precursors of HUCB-NSCs was measured with Alamar Blue assay and immunfluorescence stainings. High concentration of MeHgCl (1 µM) significantly decreased viability of cells and their ability to proliferate. The response of cells to the toxic effect of MeHgCl was different depending on the type of adhesive substrate. Domains covered with fibronectin or vitronectin, decreased significantly HUCB-NSC sensitivity to the neurotoxin when compared to poly-L-lysine. Our results suggest that receptor mediated interactions on cell membrane/biomolecule interface may be protective in neural stem cells’ response to certain neurotoxins. Supported by MSHE grant No 5978/B/P01/38 and NN 302663940
6

Immunomodulatory capacity of Wharton’s Jelly mesenchymal stem cells after stimulation in vitro: cytokines and growth factors transcriptional response analysis

71%
Lech W., Zychowicz M., Figiel-Dabrowska A., Domanska-Janik K., Buzanska L.
Acta Neurobiologiae Experimentalis
|
2017
|
tom 77
|
nr Suppl.1
INTRODUCTION: Mesenchymal Stem Cells (MSC) possess ability to release cytokines and growth factors that suppress immune responses and stimulate tissue regeneration. Wharton’s Jelly-derived MSC (WJ-MSC) in the addition to the strong adjuvant properties are characterized by low immunogenicity. AIM(S): The aim of this study is to verify immunomodulatory properties of WJ‑MSC after TNFα and IFNγ stimulation in vitro, by comparative analysis of the expression of cytokines and growth factors they produce. METHOD(S): WJ-MSC isolated from human umbilical cords were cultured in closed system that provides a constant 5% oxygen concentration. We compared immunomodulatory properties of WJ-MSC in 2D or 3D structures (scaffolds) made in our laboratory. Both of those cell populations were cultured in medium with/ without stimulate factors: TNF‑α and IFN‑γ. After stimulation, 2D and 3D cell cultures were characterized with quantitative RT-PCR for the expression of various cytokines and growth factors with non-stimulated 2D cells as an internal control. WJ-MSC grown in 3D were also characterized by live/dead cells presence which were labeled with calcein AM/ethidium homodimer. RESULTS: The obtained results indicated increased expression of mRNA in 3 D structures vs. control 2 D cells for almost all analyzed cytokines: IL‑6, TGF‑β1 , BDNF, GDNF, EGF, bFGF. Moreover, TNF‑α and IFN‑γ stimulation causes even further increase of mRNA expression of those cytokines in 3 D cultures compared to non-stimulated 2 D control. In our scaffolds models, the intercellular connections which were labeled in live cells with calcein AM have been observed already after 24h of culture and are visible also during the next days of analysis. CONCLUSIONS: Finally we can conclude, that WJ-MSC produce immunomodulatory factors, and their expression can be modulated by stimulation with chosen cytokines and 3D microenvironment. Such properties of WJ-MSC are important for the potential therapeutic application in the treatment of the diseases of inflammatory and autoimmune origin. FINANCIAL SUPPORT: The work was supported by National Centre for Research and Development grant No STRATEGMED1/234261/2/NCBR/2014.
7

Lowered oxygen concentration and culture in 3-dimension structure enhance immunological response of WJ-MSC to proinflammatory factors

71%
Figiel-Dabrowska A., Lech W., Zychowicz M., Domanska-Janik K., Sarnowska A.
Acta Neurobiologiae Experimentalis
|
2017
|
tom 77
|
nr Suppl.1
INTRODUCTION: MSC-based therapy is becoming more and more common treatment of various diseases, albeit still as an experimental approach. According to present‑day literature, the therapeutic effects of transplanted cells would not be ascribed to their differentiation, trans‑differentiation or repopulation but rather to their paracrine effect on damaged tissue. This way of treatment can be initiated and enhanced by local environmental mediators. AIM(S): The aim of this study was to assess the influence of inflammation specific environment in vitro on secretory WJ-MSC properties and to evaluate the possibility of programmed and controlled induction and enhancement of anti‑inflammatory cell properties in the context of further cell therapy. METHOD(S): Our experiments were based on reconstruction in vitro the environment to which therapeutic cells (WJ‑MSC) are usually transplanted. The inflammatory conditions that occur around the transplant were reproduced through TNFα and IFNγ stimulation. Tissue specific oxygen concentration (5%), 3‑dimension transplant structure and chemical composition of the indirect transplanted cell surrounding as determined by additional scaffold ingredients (fibrin and platelet lysate) were also reconstituted. RESULTS: Carried experiments have shown specific changes in the secreted cytokine pallet induced in vitro by the inflammation‑like WJ‑MSC surrounding. We have proved that environmental modifications cause changes in synthesis and secretion of the determined proteins. Both, the physioxia introduced in our in vitro experiments and WJ cells cultured in 3-dimensional structures enhanced cytoprotective paracrine properties of WJ-MSC. Additional reinforcing effect was observed when therapeutic cells were transplanted on platelet lysate – containing scaffolds. CONCLUSIONS: Presented results indicate that by optimization of cell culture and transplantation conditions we could control and enhance cytokine-connected therapeutic properties of MSC. FINANCIAL SUPPORT: The work was supported by National Centre for Research and Development grant No Strategmed 1/234261/2/NCBR/2014.
8

Reprogramming of somatic cells: possible methods to derive safe, clinical-grade human induced pluripotent stem cells

71%
Augustyniak J., Zychowicz M., Podobinska M., Barta T., Buzanska L.
Acta Neurobiologiae Experimentalis
|
2014
|
tom 74
|
nr 4
Derivation of pluripotent stem cells from adult somatic tissues by reprogramming technology has opened new therapeutic possibilities. Current most efficient procedures for derivation of induced pluripotent stem (iPS) cells are based on the viral vectors, which represent the danger of insertional mutagenesis during incorporation of introduced genes into the host genome. To circumvent this problem, the new, safe, non-integrative and non-viral strategies of reprogramming have been developed. In this review we discuss novel DNA-free and viral-free methods of reprogramming to iPS cells including protein transduction, mRNA and microRNA delivery.
9

Patterning of human cord blood-derived stem cells on single cell posts and lines: implications for neural commitment

61%
Zychowicz M., Mehn D., Ruiz A., Frontczak-Baniewicz M., Rossi F., Buzanska L.
Acta Neurobiologiae Experimentalis
|
2012
|
tom 72
|
nr 4
Using stem cells (SC) in new strategies for clinical treatment requires control of stem cell fate decision and the ability to govern their patterning and commitment in tissue engineering. Neural stem cells and other adult SC can respond to the different components of the microenvironment and their spatial arrangement in the stem cell niche. It has been shown previously by our group that different composition and architecture of patterned bioactive domains influence the developmental response of neural stem cells. In the present report we verify the commitment and differentiation of neural stem cells derived from human cord blood (HUCB-NSC) by a single cell patterning system. Microcontact printing technology was used to generate single cell positioning areas of different geometry: 10 ^m-thin lines and 10 ^m-width one cell posts. The commitment and differentiation of HUCB-NSC cells cultured on different surfaces were dependent on the geometry and the type of biomateriał present in bioactive domains. Fibronectin promoted neuronal protrusion outgrowth (P-tubulin III and MAP-2 positive cells) and gap junction development (Cx43 marker) between cells growing on lines while poly-L-lysine promoted HUCB-NSC differentiation into astrocytic, glial fibrillary acidic protein expressing (GFAP positive) cell phenotype. Here we also demonstrate by scanning electron microscopy that morphology of cells on the patterned surface is highly dependent upon the type of biomolecules used for printing. These kinds of platforms can be used for investigating the influence of spatial organization of environment on the SC fate decision and for studying the molecular processes occurring in a single cell.
10

Immune response following grafting of human umbilical cord blood neural stem cells (HUCB-NSC) into adult rats with ouabain (OUA) induced brain lesion

61%
Jablonska A., Kozlowska H., Zychowicz M., Wanacka E., Domanska-Janik K., Lukomska B.
Acta Neurobiologiae Experimentalis
|
2009
|
tom 69
|
nr 1
Many types of neural progenitors from different sources have been tested for experimental therapy in CNS injuries. We have established neural stem cell line from human cord blood (HUCB-NSC). In vitro evidence has suggested that HUCB-NSC are not immunogenic however their transplantation (tx) into adult rats led to the graft cell rejection. The question arises what is the nature of the host immune response to transplanted HUCB-NSC? Methods: 2 × 104 HUCBNSC were tx into corpus callosum of a focal brain injury induced by OUA injection (1 μl/5 mmol) into striatum of adult Wistar rats. At 1, 3, 7 and 14 days thereafter brains were removed and immunocytochemical analysis for T cells (CD5), B cells (CD45), macrophages (ED1) and neutrophils (CD15) was performed. Results: One day after HUCB-NSC tx, most cells remained in the injection site and only few cells migrated to the lesion area. Concomitantly, infi ltration of ED1+ and CD15+ cells with occasional appearance of CD5+ and CD45+ cells was seen around the graft and close to the lesion. At 3rd day after HUCB-NSC tx, the infi ltrate of ED1+ cells increased however, the number of CD15+, CD5+ and CD45+ cells stayed unchanged. At 7th day after HUCB-NSC tx, the number of ED1+ and CD15+ cells was reduced dramatically compare to the 3rd day and only single CD5+ and CD45+ cells were observed. By 14 days, limited number of ED1+, CD15+, CD5+ and CD45+ cells were found most likely due to the scar formation and rejection of HUCB-NSC. At that time no viable HUCB-NSC have been noticed in brain tissue of the host Conclusion: Transfer of HUCB-NSC into ouabain induced brain lesion rats elicits innate (macrophages/neutrophils) and adaptive (T/B lymphocytes) immune response in the acute phase post-transplantation. Supported by MSHE grant No 142/P01/2008/35
11

Controlling of HUCB-NSC culture by micropatterned biofunctionized surfaces

61%
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.
12

Protection of neural stem cells from MeHgCl effect at different biofunctonal domains

51%
Kozlowska H., Zychowicz M., Mehn D., Dziedzicka D., Nowak J., Stepien P., Rossi F., Buzanska L.
Acta Neurobiologiae Experimentalis
|
2011
|
tom 71
|
nr S
The methylmercury chloride (MeHgCl) is known to cause developmental neurotoxicity in humans. This neurotoxic reagent can induce cell death due to several cellular mechanisms including phosphorylation dependent pathways and disruption of microtubule assembly. The non transformed neural stem cell line obtained from human cord blood (HUCB-NSC) has been previously shown to be susceptible to MeHgCl in developmentally dependent manner. In this report we are trying to find out whether developmental sensitivity of HUCBNSC to MeHgCl depends upon the type of adhesive biomolecules in functional domains. Cell growth platforms microspotted with fibronectin, vitronectin and poly-L-lysine have been used to compare differentiation potential of HUCB-NSC into neuronal or astrocytic cells at various MeHgCl concentrations ranging from 0.05 to 1 μM. Developmental decisions of HUCB-NSC whether to differentiate into neuronal or glial lineage were observed at non cytotoxic concentrations of MeHgCl and were dependent on the type of bioactive domain. Generally, adhesive domains protected HUCB-NSC from cytotoxic effect of MeHgCl, since on plastic surface even the lowest concentration of toxicant (0.05 μM) significantly diminish the cell number after 48 h of incubation time as shown by Alamar Blue assay. The same tendency was observed in the proliferation response as shown by Ki67 presence or BrdU incorporation. Supported by MSHE grant No. 2211/B/ P01/2010/38 and No. 5978/B/P01/2010/38, and European Commission Joint Research Centre NanoBioscience Action.
13

Proliferation capacity of cord blood derived neural stem cell line on different micro-scale biofunctional domains

51%
Zychowicz M., Mehn D., Ruiz A., Colpo P., Rossi F., Frontczak-Baniewicz M., Domanska-Janik K., Buzanska L.
Acta Neurobiologiae Experimentalis
|
2011
|
tom 71
|
nr 1
Physical interactions of cells with the adhesive substrates of the microenvironment as well as the presence of the soluble growth factors are important for the proliferation capacity of neural stem cells. We have used biofunctionalized surface domains microcontact printed with either synthetic polyaminoacid poly-L-lysine or extracellular matrix (ECM) component such as fibronectin, to study the proliferation capacity of human umbilical cord blood-derived neural stem cells (HUCB- NSC). The proliferation measured by the expression of Ki-67 protein was accompanied by the investigation of the cell morphology under the transmission and scanning electron microscopy in different culture time, plating densities of cells and medium condition (serum-free or 2% of FBS). The poly-L-lysine domains of defined micro-scale area promoted the presence of round, loosely attached Ki-67-positive cells, while fibronectin domains of the same size allowed appearance of flattened, strongly attached cells with more differentiated phenotype. These results were in agreement with the non-specific, electrostatic type of interaction between cell and substrate on poly-L-lysine and integrin receptor-mediated specific adhesion on fibronectin. In this report we have described in vitro culture conditions, which allow for immobilization of the non- differentiated and highly proliferating population of neural stem/progenitor cells to the biofunctionalized surface. The microarrays with bioactive domains allocating non-differentiated and proliferating neural stem/progenitor cells may find application for drug and chemicals toxicology screening of diverse factors influencing neural development.
14

Human cord blood cells in neurotoxicology: Advantage of emerging technologies

51%
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.
15

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

51%
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.
16

Analysis of the neurogenic potential of human umbilical cord blood neural stem cells (HUCB-NSC) after their transplantation into rat brain

51%
Kozlowska H., Markiewicz I., Jablonska A., Habich A., Zychowicz M., Wanacka E., Domanska-Janik K., Lukomska B.
Acta Neurobiologiae Experimentalis
|
2009
|
tom 69
|
nr 1
Transplantation (tx) of neural stem cells (NSC) is the key strategy of cell replacement therapy in the central nervous system. The goal of the study was to compare survival, migration and differentiation of HUCB-NSC after their tx into the brain of neonatal and adult Wistar rats. Methods: HUCB-NSC (2 × 104 ) labeled with CMFDA were tx into SVZ of the postnatal day 0 (P0) rats or into intact brain of adult rats. After 1, 3, 7, 14 or 21 days brains were removed, frozen and cut into 20 μm coronary slices, then immunohistochemical studies were performed to visualize HUCB-NSC fate in the brain. Results: In neonatal rats, 3 days after tx most of HUCB-NSC remained in the graft. During the fi rst week HUCB-NSC started to disperse and migrate. HUCB-NSC situated at the periphery of the graft or in migratory stream display proliferation marker (Ki67). After 7ñ21 days HUCB-NSC survived in the host brain with many cells expressing neuronal or astrocytic phenotypes. Few of HUCB-NSC presented the features of adult neurons (MAP2+ with long protrusions). In adult rats, 3 days after tx, HUCB-NSC form dense deposit with single cells migrating into brain tissue. Most of the HUCB-NSCs stayed undifferentiated with few cells expressing neuronal (NF200) or astrocytic (GFAP) markers. After 7 days numerous HUCB-NSC situated inside the graft underwent degeneration and subsequent depletion. Tx HUCB-NSC induced infl ammatory response detected by macrophage/microglia (ED1+) accumulation and astrogliosis (GFAP+). No viable HUCB-NSC were found after 14 days. Conclusions: Host environment dictates the fate of tx neural stem cells derived from human cord blood however immunological response in adult rats limits the time of observation due to short survival of HUCB-NSC. Supported by MSHE grant no 2PO5A05430
17

HUCB-NSCS growth control by biofunctionalized surfaces

42%
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.
Pierwsza strona wyników Pięć stron wyników wstecz Poprzednia strona wyników Strona / 1 Następna strona wyników Pięć stron wyników wprzód Ostatnia strona wyników
JavaScript jest wyłączony w Twojej przeglądarce internetowej. Włącz go, a następnie odśwież stronę, aby móc w pełni z niej korzystać.