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1.
Sci Rep ; 10(1): 5379, 2020 03 25.
Article in English | MEDLINE | ID: mdl-32214151

ABSTRACT

Clear cell renal cell carcinoma (ccRCC) is the most common kidney cancer. Prognosis for ccRCC is generally poor since it is largely resistant to chemo- and radiotherapy. Many studies suggested that cancer stem cells/tumor initiating cells (CSCs/TICs) are responsible for development of tumor, disease progression, aggressiveness, metastasis and drug resistance. However, tumorigenic potential of CSCs/TICs isolated from established RCC cell lines - basic ccRCC research model - has never been investigated in vivo. CD105+, CD105-, CD44+ and CD44- as well as CD44-/CD105- CD44+/CD105+ and CD44-/CD105+ cells were isolated from Caki-1 RCC cell line, confirming coexistence of multiple subpopulations of stem-related phenotype in stable cell line. Sorted cells were injected subcutaneously into NOD SCID mice and tumor growth was monitored with MRI and PET/CT. Tumor growth was observed after implantation of CD105+, CD44+, CD44-, CD44-/CD105+ and CD44-/CD105- but not CD105- or CD44+/CD105+. Implantation of CD44-/CD105- cells induced tumors that were characterized by longer T1 and distinct metabolic pattern than other tumors. All the tumors were characterized by low uptake of [18F]FDG. CD105+ and CD44- tumors expresses Nanog and Oct-4, while CD44- tumors additionally expressed endothelial cell marker - CD31.


Subject(s)
Carcinoma, Renal Cell/immunology , Endoglin/immunology , Hyaluronan Receptors/immunology , Animals , Biomarkers, Tumor/immunology , Biomarkers, Tumor/metabolism , Carcinoma, Renal Cell/pathology , Cell Line, Tumor , Cell Separation , Endoglin/metabolism , Humans , Hyaluronan Receptors/metabolism , Kidney Neoplasms/immunology , Kidney Neoplasms/pathology , Male , Mice , Mice, Inbred NOD , Mice, SCID , Neoplastic Stem Cells/immunology , Neoplastic Stem Cells/metabolism , Positron Emission Tomography Computed Tomography , Prognosis
2.
Sci Rep ; 9(1): 799, 2019 01 28.
Article in English | MEDLINE | ID: mdl-30692571

ABSTRACT

The most recent research concerning amyotrophic lateral sclerosis (ALS) emphasizes the role of glia in disease development. Thus, one can suspect that the effective therapeutic strategy in treatment of ALS would be replacement of defective glia. One of the basic problems with human glial progenitors (hGRPs) replacement strategies is the time needed for the cells to become fully functional in vivo. The lifespan of most popular high copy number SOD1 mutant mice might be too short to acknowledge benefits of transplanted cells. We focused on developing immunodeficient rag2-/- model of ALS with lower number of transgene copies and longer lifespan. The obtained hSOD1/rag2 double mutant mice have been characterized. QPCR analysis revealed that copy number of hSOD1 transgene varied in our colony (4-8 copies). The difference in transgene copy number may be translated to significant impact on the lifespan. The death of long- and short-living hSOD1/rag2 mice is preceded by muscular weakness as early as one month before death. Importantly, based on magnetic resonance imaging we identified that mutant mice demonstrated abnormalities within the medullar motor nuclei. To conclude, we developed long-living double mutant hSOD1/rag2 mice, which could be a promising model for testing therapeutic utility of human stem cells.


Subject(s)
Amyotrophic Lateral Sclerosis/diagnostic imaging , DNA Copy Number Variations , DNA-Binding Proteins/genetics , Superoxide Dismutase-1/genetics , Amyotrophic Lateral Sclerosis/genetics , Amyotrophic Lateral Sclerosis/immunology , Amyotrophic Lateral Sclerosis/physiopathology , Animals , DNA-Binding Proteins/metabolism , Disease Models, Animal , Disease Progression , Female , Gene Knockout Techniques , Humans , Immunocompromised Host , Male , Mice , Mice, Transgenic , Protein Folding , Severity of Illness Index , Spinal Cord/diagnostic imaging , Spinal Cord/metabolism , Superoxide Dismutase-1/chemistry , Superoxide Dismutase-1/metabolism , Trigeminal Motor Nucleus/diagnostic imaging , Trigeminal Motor Nucleus/metabolism
3.
Br J Ophthalmol ; 92(8): 1142-7, 2008 Aug.
Article in English | MEDLINE | ID: mdl-18653608

ABSTRACT

BACKGROUND/AIMS: To evaluate the retinal toxicity of Brilliant Blue G (BBG) following intravitreal injection in rat eyes and examine the biocompatibility and the staining properties in humans. METHODS: BBG was injected into the 11 rat eyes to evaluate toxic effects with balanced salt solution (BSS) serving as control. Retinal toxicity was assessed by retinal ganglion cell (RGC) counts and by light microscopy 7 days later. In addition, BBG was applied during vitrectomy for macular hole (MH) (n = 15) or epiretinal membranes (ERM) (n = 3) in a prospective, non-comparative consecutive series of patients. Before and after surgery, all patients underwent a complete clinical examination including measurement of best corrected visual acuity (VA) and intraocular pressure, perimetry, fundus photography and optical coherence tomography. Patients were seen 1 day before surgery and then in approximately four weeks intervals. RESULTS: No significant reduction in RGC numbers and no morphological alterations were noted. A sufficient staining of the internal limiting membrane (ILM) was seen in patients with MH, while the staining pattern in ERM cases was patchy, indicating that parts of the ILM were peeled off along with the ERM in a variable extent. All MHs could be closed successfully. VA improved in 10 eyes (56%; 8/15 MH patients, 2/3 ERM patients), was unchanged in four eyes (22%; all MH patients) and was reduced in four eyes (22%; 3/15 MH, 1/3 ERM). No toxic effects attributable to the dye were noted during patient follow-up. The ultrastructure of tissue harvested during surgery was unremarkable. CONCLUSION: Brilliant Blue provides a sufficient and selective staining of the ILM. No retinal toxicity or adverse effects related to the dye were observed in animal and human studies. The long-term safety of this novel dye will have to be evaluated in larger patient series and a longer follow-up.


Subject(s)
Benzenesulfonates/toxicity , Coloring Agents/toxicity , Retina/drug effects , Aged , Animals , Cell Count , Epiretinal Membrane/diagnosis , Epiretinal Membrane/pathology , Epiretinal Membrane/surgery , Female , Humans , Male , Middle Aged , Prospective Studies , Rats , Rats, Inbred BN , Retina/pathology , Retina/ultrastructure , Retinal Ganglion Cells/drug effects , Retinal Ganglion Cells/pathology , Retinal Perforations/surgery , Staining and Labeling/methods , Vitrectomy/methods
4.
Photochem Photobiol ; 65(5): 855-61, 1997 May.
Article in English | MEDLINE | ID: mdl-9155257

ABSTRACT

The photodynamic effect of Victoria blue BO (VB-BO) and photoirradiation on peripheral blood mononuclear cells was studied. The cells were preincubated with VB-BO followed by photoirradiation and overnight culture. The highest percentage of dead cells (propidium iodide assay in flow cytometry) was seen in the monocyte population. The lymphocytes showed a lower sensitivity to VB-BO photodynamic action than the monocytes (12% vs 80% of PI-positive cells). The effect of VB-BO and phototreatment on lymphocyte function was studied using a mitogen-induced proliferation assay. A decrease of mitogen response was observed. The VB-BO and photoirradiation were also used on leukemic cells. The leukemic cells from acute myeloid leukemia and B precursors leukemia were sensitive to VB-BO photodynamic action. The high VB-BO sensitivity of monocytes and leukemic cells (myeloid and lymphoid B derived) suggests possible application of VB-BO for selective depletion of monocytes or sensitive leukemic cells.


Subject(s)
Antineoplastic Agents/pharmacology , Leukemia/pathology , Leukocytes, Mononuclear/drug effects , Photosensitizing Agents/pharmacology , Quaternary Ammonium Compounds/pharmacology , Antiviral Agents/pharmacology , Bone Marrow Cells , Cell Survival/drug effects , Cell Survival/radiation effects , Fluorescent Dyes , Humans , In Vitro Techniques , Leukocytes, Mononuclear/radiation effects , Monocytes/drug effects , Monocytes/radiation effects , Photochemistry , Pyrimidinones/pharmacology , Radiation Tolerance/drug effects
5.
Photochem Photobiol ; 58(3): 356-61, 1993 Sep.
Article in English | MEDLINE | ID: mdl-8234469

ABSTRACT

Photodynamic induced cytotoxicity by Victoria blue BO (VB-BO), merocyanine 540 (MC540), Nile blue A (NB) and 4-tetrasulfonatophenyl-porphyrin (4-TSPP) has been studied on two human leukemic cell lines: K-562 and TF-1. Cells were incubated with dyes and irradiated with different doses of white light. Cell survival was assessed by propidium iodide (PI) staining using flow cytometry analysis. Concentrations of 5 x 10(-8) M VB-BO were found to kill 75% of cells, and a concentration of 1 x 10(-7) M induced more than 99% of cell killing. To obtain the same cytotoxic level, the presence of 2.6 x 10(-5) M of MC540 during irradiation was needed. Under the conditions used, NB was ineffective as a photosensitizer, although uptake studies showed that this dye was taken by the cells in much greater amounts than any other studied dye. Cell cycle distribution of TF-1 cells, surviving MC540 or VB-BO photosensitization has been studied by flow cytometry analysis after staining with Hoechst 33342 and PI. It was found that cells in G1 phase were slightly more resistant toward MC540- and VB-BO-mediated photosensitization than cells in other phases of the cell cycle.


Subject(s)
Antineoplastic Agents/pharmacology , Photochemotherapy , Quaternary Ammonium Compounds/pharmacology , Cell Cycle/drug effects , Drug Screening Assays, Antitumor , Humans , Leukemia , Oxazines/pharmacology , Photosensitizing Agents/pharmacology , Porphyrins/pharmacology , Pyrimidinones/pharmacology , Tumor Cells, Cultured
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