Natalia Gurtowska

Adipose-Derived Stem Cells as a Tool in Cell-Based Therapies

Recent development in stem cell isolation methods and expansion under laboratory conditions create an opportunity to use those aforementioned cells in tissue engineering and regenerative medicine. Particular attention is drawn towards mesenchymal stem cells (MSCs) being multipotent progenitors exhibiting several unique characteristics, including high proliferation potential, self-renewal abilities and multilineage differentiation into cells of mesodermal and non-mesodermal origin. High abundance of MSCs found in adipose tissue makes it a very attractive source of adult stem cells for further use in regenerative medicine applications. Despite immunomodulating properties of adipose-derived stem cells (ASCs) and a secretion of a wide variety of paracrine factors that facilitate tissue regeneration, effectiveness of stem cell therapy was not supported by the results of clinical trials. Lack of a single, universal stem cell marker, patient-to-patient variability, heterogeneity of ASC population combined with multiple widely different protocols of cell isolation and expansion hinder the ability to precisely identify and analyze biological properties of stem cells. The above issues contribute to conflicting data reported in literature. We will review the comprehensive information concerning characteristic features of ASCs. We will also review the regenerative potential and clinical application based on various clinical trials.

Does aging of mesenchymal stem cells limit their potential application in clinical practice

Mesenchymal stem cells (MSCs) are in the center of attention of many investigators due to easy isolation from many tissues. MSC capability to differentiate into many cell types makes them a starting point of many new therapies, especially in tissue engineering. However, understanding the process of MSC aging is crucial for selecting donors for cellular therapies, which is necessary for successful treatment. Cell changes can be divided into three major groups. Changes which affect their proliferate rate, differentiation capability and genome stability lead to decrease of their usefulness in new therapies. There are many tools that can be used to describe and measure some features of aging in MSCs but the essence of this process is still unclear. The aim of this review is to take a deep look into the influence of donor age and in vitro aging on MSC properties.

Ciprofloxacin is a potential topoisomerase II inhibitor for the treatment of NSCLC.

Lung cancer is one of the most common tumors and its treatment is still inefficient. In our previous work we proved that ciprofloxacin has a different influence on five cancer cell lines. Here, we aimed to compare the biological effect of ciprofloxacin on cell lines representing different responses after treatment, thus A549 was chosen as a sensitive model, C6 and B16 as highly resistant. Three different cell lines were analyzed (A549, B16 and C6). The characterization of continuous cell growth was analyzed with the Real-Time Cell Analyzer (RTCA)-DP system. Cytoskeletal changes were demonstrated using immunofluorescence. The cell cycle was analyzed using flow cytometry. Ciprofloxacin was cytostatic only against the A549 cell line. In the case of other tested cell lines a cytostatic effect was not observed. Cytoskeletal analysis confirms the results obtained with RTCA-DP. A549 cells were inhibited in the G2/M phase suggesting a mechanism related to topoisomerase II inhibition. The biological effects of ciprofloxacin support the hypothesis that this drug can serve as an adjuvant treatment for lung cancer, due to its properties enabling topoisomerase II inhibition.

Does ciprofloxacin have an obverse and a reverse

Ciprofloxacin is an antibiotic that belongs to fluoroquinoles, characterized by broad spectrum of action against pathogens, especially Gram(-) aerobic bacilli. For a long time, it has been thought that ciprofloxacin has an effect only on bacterial cells. Now it is known, that this drug can significantly affect eukaryotic cells including human cancer cells. Its bactericidal action relay on inhibition of topoisomerase II, enzyme responsible for alterations in 3D structure of DNA during replication, transcription and chromatin condensation. Thanks to that, ciprofloxacin can induce cell cycle arrest and apoptosis of cancer cells. The effectiveness of ciprofloxacin was confirmed in several in vitro studies on tumor cell lines such as: human bladder cells, leukaemic cell lines, human osteosarcoma cells, human prostate cancer cells, human colorectal carcinoma cells and human non-small cell lung cancer cell line. Ciprofloxacin is particularly effective against non-small cell lung cancer mainly due to accumulation of ciprofloxacin in lung tissue after intravenous administration and its toxicity against lung cancer lines in vitro in a concentration and time-dependent manner.

Does the liposuction method influence the phenotypic characteristic of human adipose-derived stem cells?

Statistical analysis revealed significant differences in antigen expression of 58 markers of the 242 studied. The method of liposuction has no significant impact on antigens profile in cultured ASCs (adipose-derived stem cells).

How to isolate urothelial cells? Comparison of four different methods and literature review

The aim of this study is to present the comparison of four different methods for urothelial cell isolation and culture and compare them to methods cited in the literature. Four different techniques were examined for urothelium isolation from rat bladders. Isolation effectiveness was calculated using trypan blue assay. Confirmation of isolated cell phenotype and comparison with native bladder tissue was confirmed using immunohistochemical (IHC), immunocytochemical (ICC) and immunofluorescence (IF) analysis. The method with bladder inversion and collagenase P digestion resulted in the highest number of isolated cells. These cells showed positive expression of cytokeratin 7, 8, 18, α6-integrin and p63. Our results and the literature review showed that the best method for urothelium bladder isolation is dissection of the epithelium layer from other bladder parts and digestion of mechanically prepared tissue in a collagenase solution.

Does the Harvesting Technique Affect the Properties of Adipose-Derived Stem Cells? - The Comparative Biological Characterization.

The objective of this study was to evaluate complex biological properties of human stem cells isolated from adipose tissue (ASCs) harvested utilizing different methods: surgical resection (R), power‐assisted liposuction (PAL), and laser‐assisted liposuction (LAL). ASCs were isolated from healthy donors, due to surgical resection, power‐, and laser‐assisted liposuction. Isolated cells were characterized by their clonogenicity, proliferation rate, doubling time, multilineage differentiation, and senescence potential. The average number of ASCs from 1g/1 ml of solid adipose tissue/lipoaspirate was 2.9 × 105 ± 2.4 × 105, 1.1 × 105 ± 0.8 × 105, and 1.2 × 105 ± 0.7 × 105, respectively, for ASCsR, ASCsPAL, and ASCsLAL. However, number of colonies formed by ASCsR and ASCsPAL was significantly higher compared to the average number of colonies formed by ASCsLAL. Also, in comparison to other analyzed cell groups, ASCsPAL obtained the highest proliferative activity. All analyzed cells were characterized by stable expression of CD90 and CD44 markers during prolonged culture. Expression of CD34 and CD45 markers was decreasing in subsequent passages. Presented study shows that different ASCs collection method affects some basic characteristics of these cells, such as number of isolated cells, clonogeneity, or doubling time. J. Cell. Biochem. 118: 1097–1107, 2017.

Ciprofloxacin as a prophylactic agent against prostate cancer: A “two hit” hypothesis

More evidence indicate that prostate inflammation can lead to prostate cancer development. Prostate cancer affects elderly men. Prostate cancer prophylaxis is an important issue because life expectancy is very long now. Ciprofloxacin is an antibacterial agent used mainly in urinary tract infections and prostate inflammation. This drug acts also against cancer cells by the inhibition of topoisomerase II. These properties should allow it to inhibit the development of prostate cancer. Firstly, ciprofloxacin can stop the acute and chronic prostate inflammation which can lead to cancer development. Secondly, ciprofloxacin can potentially kill prostate cancer cells in their early stage of development. Ciprofloxacin accumulates mainly in the prostate after oral intake thus ciprofloxacin seems to be a perfect candidate as a prophylactic agent.