Wyniki wyszukiwania - Biblioteka Nauki

1

Optimisation of transfection conditions of CD34+ hematopoietic cells derived from human umbilical cord blood

100%

Ołdak T. , Kruszewski M. , Machaj E. K. , Gajkowska A. , Pojda Z.

Acta Biochimica Polonica

|

2002

|

tom 49

|

nr 3

625-632

EN

Human umbilical cord blood is frequently used as a source of transplantable hematopoietic cells and more recently as a target of gene therapy - a new approach for treatment of various disorders. The aim of our study was optimisation of the transfection conditions of cord blood-derived CD34+ hematopoietic cells. Mononuclear cells fraction was isolated from cord blood samples by density gradient centrifugation. Subsequently, CD34+ hematopoietic cells were separated on immunomagnetic MiniMACS columns. Pure population of CD34+ cells was incubated in a serum free medium supplemented with thrombopoietin, stem cell factor and Flt-3 ligand for 48 h and then transfected with plasmid DNA carrying the enhanced version of green fluorescent protein (EGFP) as a reporter gene. We studied the influence of various pulse settings and DNA concentrations on the transfection efficiency, measured by flow cytometry as the fluorescence of target cells due to the expression of EGFP. The optimal settings were as follows: 4 mm cuvette, 1600 μF, 550 V/cm, and 10 μg of DNA per 500 μl. With these settings we obtained a high transfection frequency (41.2%) without a marked decrease of cell viability. An increase of the pulse capacitance and/or of DNA concentration resulted in a greater electroporation efficiency, but also in a decrease of cell viability. In conclusion, the results described here allow one to recommend electroporation as an efficient method of gene delivery into CD34+ hematopoietic cells derived from human umbilical cord blood.

2

International Symposium on Umbilical Cord Blood Transplantation: new insights and future directions

88%

Smogorzewska E. M.

Archivum Immunologiae et Therapiae Experimentalis

|

2003

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tom 51

|

nr 5

3

Comparison of gene expression of mitogenic kinin path in adherent and non-adherent CD 34-stem cells using oligonucleotide microarrays

75%

Stojko R. , Witek A. , Glogowska J. , Mazurek U. , Chromy G. , Wilk K. , Witek L. , Bojdys-Szyndlar M. , Machaj K. , Pojda Z.

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tom 46

|

nr 1

45-50

4

Nonviral transfection of human umbilical cord blood dendritic cells is feasible, but the yield of dendritic cells with transgene expression limits the application of this method in cancer immunotherapy

75%

Markowicz S. , Niedzielska J. , Kruszewski M. , Ołdak T. , Gajkowska A. , Machaj E. K. , Skurzak H. , Pojda Z.

Acta Biochimica Polonica

|

2006

|

tom 53

|

nr 1

203-212

EN

Dendritic cells (DC) generated from human umbilical cord blood might replace patients' DC in attempts to elicit tumor-specific immune response in cancer patients. We studied the efficiency of transfection of human cord blood DC with plasmid DNA carrying the enhanced version of green fluorescent protein (EGFP) as a reporter gene, to test if nonviral gene transfer would be a method to load DC with protein antigens for immunotherapy purposes. Cord blood mononuclear cells were cultured in serum-free medium in the presence of granulocyte-monocyte colony stimulating factor (GM-CSF), stem cell factor (SCF) and Flt-3 ligand (FL), to generate DC from their precursors, and thereafter transfected by electroporation. Maturation of DC was induced by stimulation with GM-CSF, SCF, FL and phorbol myristate acetate (PMA). Transfected DC strongly expressed EGFP, but transfection efficiency of DC, defined as HLA-DR+ cells lacking lineage-specific markers, did not exceed 2.5%. Expression of the reporter gene was also demonstrated in the DC generated from transfected, purified CD34+ cord blood cells, by stimulation with GM-CSF, SCF, FL, and tumor necrosis factor α (TNF-α). Transfection of CD34+ cells was very efficient, but proliferation of the transfected cells was much reduced as compared to the untransfected cells. Therefore, the yield of transgene-expressing DC was relatively low. In conclusion, nonviral transfection of cord blood DC proved feasible, but considering the requirements for immunotherapy in cancer patients, transfection of differentiated DC or generation of DC from transfected hematopoietic stem cells provide only a limited number of DC expressing the transgene.

5

Krew pępowinowa: pozyskiwanie, preparatyka, przechowywanie

75%

Zagórska-Szlufik P.

Laboratorium Medyczne

|

2020

|

tom nr 1

20--25

PL

Krew pępowinowa jest źródłem hematopoetycznych komórek macierzystych. Zanim stwierdzono jej przydatność w medycynie, była traktowana jako odpad medyczny i wraz z łożyskiem trafiała do utylizacji. Ze względu na unikatowe właściwości, komórki macierzyste izolowane z krwi pępowinowej znajdują coraz większe zastosowanie w terapiach. Obecnie z ich wykorzystaniem leczy się ponad 70 różnych schorzeń, m.in.: nowotwory hematologiczne, nienowotworowe choroby układu krwiotwórczego, choroby autoimmunologiczne i dziedziczne. Proces pozyskania krwi pępowinowej jest bezpieczny zarówno dla matki, jak i dziecka. Niniejsze opracowanie opisuje proces pozyskania krwi pępowinowej oraz jej preparatyki i przechowywania na podstawie działalności Publicznego Banku Komórek Macierzystych Regionalnego Centrum Naukowo-Technologicznego.

EN

Umbilical cord blood is a source of haematopoietic stem cells. Before this biospecimen has been proven to be useful in medicine, it was treated as medical waste and disposed of along with the placenta. Due to their unique properties, stem cells isolated from umbilical cord blood are being increasingly widely applied in therapies. Currently, they are used to treat over 70 different diseases, e.g. haematological cancers, non-cancerous haematopoietic diseases, autoimmune, and hereditary diseases. The process of obtaining umbilical cord blood is safe for both mother and child. This paper describes the process of collecting umbilical cord blood as well as its preparation and storage based on the operations of the Public Stem Cell Bank of the Regional Science and Technology Centre.

6

Detection of a CD4plusCD8minusCD3minus cell subpopulation during the differentiation of cord blood CD34plus cells into T cells in vitro

63%

Jin J. G. , Bai B. J. , Yao Z. J. , Wu R. N. , Feng K. , Hu J. W. , Hu L. D. , Jiang M. , Liao L. , Chen H.

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tom 57

|

nr 3

213-219

7

The influence of BDNF on human umbilical cord blood stem/progenitor cells: Implications for stem cell-based therapy of neurodegenerative disorders

63%

Paczkowska E. , Luczkowska K. , Piecyk K. , Roginska D. , Pius-Sadowska E. , Ustianowski P. , Cesarska E. , Dolegowska B. , Celewicz Z. , Machalinski B.

Acta Neurobiologiae Experimentalis

|

2015

|

tom 75

|

nr 2

EN

Umbilical cord blood (UCB)-derived stem/progenitor cells (SPCs) have demonstrated the potential to improve neurologic function in different experimental models. SPCs can survive after transplantation in the neural microenvironment and induce neuroprotection, endogenous neurogenesis by secreting a broad repertoire of trophic and immunomodulatory cytokines. In this study, the influence of brain-derived neurotrophic factor (BDNF) pre-treatment was comprehensively evaluated in a UCB-derived lineage-negative (Lin-) SPC population. UCB-derived Lin- cells were evaluated with respect to the expression of i) neuronal markers using immunofluorescence staining and ii) specific (TrkB) receptors for BDNF using flow cytometry. Next, after BDNF pre-treatment, Lin- cells were extensively assessed with respect to apoptosis using Western blotting and proliferation via BrdU incorporation. Furthermore, NT-3 expression levels in Lin- cells using RQ PCR and antioxidative enzyme activities were assessed. We demonstrated neuronal markers as well as TrkB expression in Lin- cells and the activation of the TrkB receptor by BDNF. BDNF pre-treatment diminished apoptosis in Lin- cells and influenced the proliferation of these cells. We observed significant changes in antioxidants as well as in the increased expression of NT-3 in Lin- cells following BDNF exposure. Complex global miRNA and mRNA profiling analyses using microarray technology and GSEA revealed the differential regulation of genes involved in the proliferation, gene expression, biosynthetic processes, translation, and protein targeting. Our results support the hypothesis that pre-treatment of stem/progenitor cells could be beneficial and may be used as an auxiliary strategy for improving the properties of SPCs.

8

Autologous cord blood transfusion in preterm infants - could its humoral effect be the key to control prematurity-related complications? A preliminary study

63%

Kotowski M. , Litwinska Z. , Klos P. , Pius-Sadowska E. , Zagrodnik-Ulan E. , Ustianowski P. , Rudnicki J. , Machalinski B.

Journal of Physiology and Pharmacology

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2017

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tom 68

|

nr 6

9

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

63%

Dziedzicka D. , Zychowicz M. , Stepien P. , Buzanska L.

Acta Neurobiologiae Experimentalis

|

2013

|

tom 73

|

nr 1

EN

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

10

Optimisation of transfection conditions of CD34plus hematopoietic cells derived from human umbilical cord blood

63%

Oldak T. , Kruszewski M. , Machaj E. K. , Gajkowska A. , Pojda Z.

Acta Biochimica Polonica

|

2002

|

tom 49

|

nr 3

EN

Human umbilical cord blood is frequently used as a source of transplantable hematopoietic cells and more recently as a target of gene therapy - a new approach for treatment of various disorders. The aim of our study was optimisation of the transfection conditions of cord blood-derived CD34+ hematopoietic cells. Mononuclear cells fraction was isolated from cord blood samples by density gradient centrifugation. Subsequently, CD34+ hematopoietic cells were separated on immuno- magnetic MiniMACS columns. Pure population of CD34+ cells was incubated in a serum free medium supplemented with thrombopoietin, stem cell factor and Flt-3 ligand for 48 h and then transfected with plasmid DNA carrying the enhanced version of green fluorescent protein (EGFP) as a reporter gene. We studied the influence of various pulse settings and DNA concentrations on the transfection efficiency, measured by flow cytometry as the fluorescence of target cells due to the expression of EGFP. The optimal settings were as follows: 4 mm cuvette, 1600 iF, 550 V/cm, and 10 ug of DNA per 500 ul. With these settings we obtained a high transfection frequency (41.2%) without a marked decrease of cell viability. An increase of the pulse capacitance and/or of DNA concentration resulted in a greater electroporation efficiency, but also in a decrease of cell viability. In conclusion, the results described here allow one to recommend electroporation as an efficient method of gene delivery into CD34+ hematopoietic cells derived from human umbilical cord blood.

11

Nonviral transfection of human umbilical cord blood dendritic cells is feasible, but the yield of dentritic cells with transgene expression limits the application of this method in cancer immunotherapy

63%

Markowicz S. , Niedzielska J. , Kruszewski M. , Oldak T. , Gajkowska A. , Machaj E. K. , Skurzak H. , Pojda Z.

Acta Biochimica Polonica

|

2006

|

tom 53

|

nr 1

EN

Dendritic cells (DC) generated from human umbilical cord blood might replace patients' DC in attempts to elicit tumor-specific immune response in cancer patients. We studied the efficiency of transfection of human cord blood DC with plasmid DNA carrying the enhanced version of green fluorescent protein (EGFP) as a reporter gene, to test if nonviral gene transfer would be a method to load DC with protein antigens for immunotherapy purposes. Cord blood mononuclear cells were cultured in serum-free medium in the presence of granulocyte-monocyte colony stimulating factor (GM-CSF), stem cell factor (SCF) and Flt-3 ligand (FL), to generate DC from their precursors, and thereafter transfected by electroporation. Maturation of DC was induced by stimulation with GM-CSF, SCF, FL and phorbol myristate acetate (PMA). Transfected DC strongly expressed EGFP, but transfection efficiency of DC, defined as HLA-DR+ cells lacking lineage-specific markers, did not exceed 2.5%. Expression of the reporter gene was also demonstrated in the DC generated from transfected, purified CD34+ cord blood cells, by stimulation with GM-CSF, SCF, FL, and tumor necrosis factor α (TNF-α). Transfection of CD34+ cells was very efficient, but proliferation of the transfected cells was much reduced as compared to the untransfected cells. Therefore, the yield of transgene-expressing DC was relatively low. In conclusion, nonviral transfection of cord blood DC proved feasible, but considering the requirements for immunotherapy in cancer patients, transfection of differentiated DC or generation of DC from transfected hematopoietic stem cells provide only a limited number of DC expressing the transgene.

12

Lead and cadmium in umbilical cord blood and maternal blood of women from Gdansk Region

63%

Szyszko M. , Giedrys A. , Krechniak J.

Acta Toxicologica

|

2004

|

tom 12

|

nr 1

13

Źródła komórek macierzystych krwiotworzenia dla potrzeb transplantacji

51%

Rzepecki P. , Gawroński K. , Młot B. , Oborska S. , Pielichowski W. , Waśko-Grabowska A.

Current Gynecologic Oncology

|

2011

|

tom 9

|

nr 3

186-198

EN

Bone marrow transplantation dates back to the ‘50s, but greatest progress in this therapeutic modality applied successfully in the treatment of several conditions took place mostly during the past 25 years. Among all organ transplantations, bone marrow transplants are second only to kidney transplants. In Poland this therapeutic technique also undergoes a rapid development. During the past 10 years, absolute numbers of various forms of transplantation increased 100-fold, starting from 6 procedures in 1989 to over 700 at present. Essentially, the term bone marrow transplantation (BMT), present in names of international associations and registers, is in fact a colloquialism (e.g. European Group for Blood and Marrow Transplantation). A much more appropriate term would be hematopoietic cell transplantation (HCT). This is because progenitor cells for transplantation may be obtained not only from bone marrow, but also from peripheral blood and umbilical cord blood. The term hematopoietic cell transplantation has a much broader meaning and includes: classic transplantation of bone marrow obtained at surgery, transplantation of peripheral blood stem cells (PBSCT) and umbilical cord blood-derived stem cells. All hematopoietic stem cells express on their surface the CD34+ antigen and glycosylated transmembrane protein, a member of the family of adhesion molecules. In healthy persons, expression of this antigen in bone marrow cells is at the level of 1-3%, while in peripheral blood – 0.01-0.1%, and in umbilical cord blood – 0.1-0.4%. The first source (transplantation material) of hematopoietic progenitor cells was bone marrow, while transplantation of stem cells obtained from peripheral blood and umbilical cord blood started much later (during the ‘80s of the past century).

14

Mesenchymal stem cells promote a primitive phenotype CD34+c-kit+ in human cord blood-derived hematopoietic stem cells during ex vivo expansion

51%

Rodriguez-Pardo V. , Vernot J. P.

Cellular and Molecular Biology Letters

|

2013

|

tom 18

|

nr 1

EN

The purpose of this study was to evaluate the influence of bone marrow-mesenchymal stem cells (BM-MSC) and exogenously added cytokines on the proliferation, primitive cell subpopulation maintenance (including the c-kit+ marker) and clonogenic capacity of hematopoietic stem cells (HSC). BM-MSC were collected from volunteer donors, isolated and characterized. Umbilical cord blood (UCB) samples were collected from healthy full-term deliveries. UCB-CD34+ cells were cultured in the presence or absence of BM-MSC and/or cytokines for 3 and 7 days. CD34+ cell proliferation was evaluated using the CSFE method and cell phenotype was determined by CD34, c-kit, CD33, CD38, HLA-DR, cyCD22 and cyCD3 detection. Cell clonogenic ability was also assessed. Exogenously added SCF, TPO and FLT3L increasedCD34+ cell proliferation in the presence or absence of BM-MSC, but with concomitant cell differentiation. Without any added cytokines, BM-MSC are able to increase the percentage of primitive progenitors as evaluated by c-kit expression and CFU-GEMM increase. Interestingly, this latter effect was dependent on both cell-cell interactions and secreted factors. A 7-day co-culture period will be optimal for obtaining an increased primitive HSC level. Including c-kit as a marker for primitive phenotype evaluation has shown the relevance of BM-MSC and their secreted factors on UCB-HSC stemness function. This effect could be dissociated from that of the addition of exogenous cytokines, which induced cellular differentiation instead.

15

The expression of B7-H1 and B7-H4 molecules on immature myeloid and lymphoid dendritic cells in cord blood of healthy neonates

51%

Kludka-Sternik M. , Serafin A. , Darmochwal-Kolarz D. , Radej S. , Rolinski J. , Leszczynska-Gorzelak B. , Oleszczuk J.

Folia Histochemica et Cytobiologica

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2010

|

tom 48

|

nr 4