Journal of Cellular Biotechnology - Volume 3, issue 1

Authors: Amin, Hawar Ahmad Muhammed | Arihan, Okan | Ragbetli, Murat Cetin

Article Type: Research Article

Abstract: BACKGROUND: Diethylnitrosamine is a carcinogenic chemical found in food unintendedly and also in different media such as cosmetics. Nigella sativa plant has anti-tumoral, anti-inflammatory and anti-diabetic activities. OBJECTIVE: Aim of this study is to investigate effect of thymoquinone -one of the active ingredients of Nigella sativa - on erythrocyte fragility in diethylnitrosamine administered rats. METHODS: 28 male Wistar-albino rats were divided into four groups; Control group (Saline/5 days, i.p.), thymoquinone group (4 mg/kg/5 days/p.o.), diethylnitrosamine group (Saline/5 days, i.p. and diethylnitrosamine 200 mg/kg/i.p. in two consecutive days) and diethylnitrosamine + thymoquinone group (4 mg/kg/5 days thymoquinone p.o. and …diethylnitrosamine 200 mg/kg/i.p. in two consecutive days). Erythrocyte fragility, hemoglobin and hematocrit counts were performed. RESULTS: Number of erythrocytes was increased significantly (p  < 0.05) in diethylnitrosamine administered group (9.05×106 /mm3 ) compared to control (7.44×106 /mm3 ) and thymoquinone (7.75×106 /mm3 ). Hematocrit value was significantly higher in diethylnitrosamine group (52.47%) compared to control (44.75%) and thymoquinone groups (46.21%) (p  < 0.05). Hemoglobin value was higher in the diethylnitrosamine administered groups (18.43 and 17.63 g/dL in diethylnitrosamine and thymoquinone + diethylnitrosamine groups respectively) compared to groups without diethylnitrosamine administration (15.15 and 15.87 g/dL in control and thymoquinone groups respectively) (p  < 0.05). Mch was found significantly higher in thymoquinone+diethylnitrosamine group (p  < 0.05). No significant difference in erythrocyte fragility was observed among groups. DISCUSSION AND CONCLUSION: Augmentation in erythrocyte, hemoglobin and hematocrit count due to diethylnitrosamine administration and a slight reversal due to thymoquinone administration were observed without statistical significance. Show more

Keywords: Thymoquinone, diethylnitrosamine, erythrocyte, rat, Nigella sativa

DOI: 10.3233/JCB-179008

Citation: Journal of Cellular Biotechnology, vol. 3, no. 1, pp. 1-7, 2017

Authors: Hiebl, Bernhard | Peters, Sinem | Gemeinhardt, Ole | Niehues, Stefan M. | Jung, Friedrich

Article Type: Research Article

Abstract: Cell exposure to cytotoxic substrates can cause decreased cell viability or cellular death through necrosis or apoptosis. The common characteristic of both reaction patterns (necrosis and apoptosis) is a compromised or damaged cell membrane, which results in a release of the cytoplasmatic enzyme lactate dehydrogenase (LDH) into the extracellular space. For this reason the content of this enzyme in the supernatants from cells can be used as an indicator of damaged cell membrane integrity and serve as a general means to assess cell viability by measuring plasma membrane permeability. However, cell culture media which are used to maintain and to …support cell growth commonly need serum supplementation, and sera already contain various LDH amounts, which may increase background absorbance in LDH release assays. Additionally the serum content varies depending on the cellular needs. For this reasons we tested the effect of different serum loads (inactivated fetal bovine serum [FBS]; 1–10 vol%) on the results of a commercially available LDH release assay (Cytotoxicity detection Kit, Roche) using 3T3 cells. An FBS concentration of 1 vol% was found to still maintain 3T3 cell viability, and after cell exposure to a cytotoxic substrate (surfactant) the cellular LDH release was higher than in the cells which were grown with higher serum concentrations (5 and 10 vol%). These results show that a serum content of 1 vol% is advantageous for cytotoxicity assays based on the LDH release, but due to the cellular differences in the need of serum components test results are limited to 3T3 cells. Show more

Keywords: Lactate dehydrogenase, biocompatibility, cytotoxicity, cell culture

DOI: 10.3233/JCB-179002

Citation: Journal of Cellular Biotechnology, vol. 3, no. 1, pp. 9-13, 2017

Authors: Saldanha, C. | Silva-Herdade, A.S.

Article Type: Research Article

Abstract: The aim of this mini review was to show data that will could open new biotechnical applications. Physiological properties of the erythrocytes obtained in inflammatory conditions simulated in vitro or in vivo in experimental animal models of inflammation or ex vivo by analysis performed in blood samples from patients with vascular diseases are herein presented. Those data obtained in vivo have been utilized in mathematical models of vascular blood circulation on search biomechanical properties of blood components. Behind the enlargement of scientific knowledge all data could open new questions giving support to create new therapeutic …drugs and to ameliorate apparatus for non-invasive detection of vascular circulatory diseases and further surgical interventions. Show more

Keywords: Kewords: Erythrocyte deformability, inflammation, nitric oxide

DOI: 10.3233/JCB-179003

Citation: Journal of Cellular Biotechnology, vol. 3, no. 1, pp. 15-20, 2017

Authors: Klicks, Julia | von Molitor, Elena | Ertongur-Fauth, Torsten | Rudolf, Rüdiger | Hafner, Mathias

Article Type: Research Article

Abstract: Skin fulfils a plethora of eminent physiological functions ranging from physical barrier over immunity shield to the interface mediating social interaction. Prone to several acquired and inherited diseases, skin is therefore a major target of pharmaceutical and cosmetic research. The lack of similarity between human and animal skin and rising ethical concerns in the use of animal models have driven the search for novel realistic three-dimensional skin models. This review provides a survey of contemporary skin models and compares them in terms of applicability, reliability, cost and complexity.

Keywords: Skin, phenotypic screening, 3D models, pharmaceutical research

DOI: 10.3233/JCB-179004

Citation: Journal of Cellular Biotechnology, vol. 3, no. 1, pp. 21-39, 2017

Authors: Rustamov, Valeh | Rudolf, Rüdiger | Yagublu, Vugar | Kuhn, Hella-Monika | Vitacolonna, Mario | Hafner, Mathias

Article Type: Research Article

Abstract: INTRODUCTION: In recent years, many different methods were introduced for generation of 3D cell culture. However, many currently available three-dimensional techniques are not suitable for certain cell lines and sometimes showed a lack of reproducibility. Therefore, specific protocols for cell lines are needed. In this work, we demonstrate different methods of generating 3D cell culture for SCC4 tongue cancer cell line and discuss their applicability. MATERIALS AND METHODS: Using three different methods, tumor spheroids were generated from SCC4 cells and cultured for 20 days. To investigate the influence of initial seeding density on spheroid morphology and size during …long term culture, the same set of different cell numbers was used for each method. Using phase-contrast microscopy, spheroids were monitored until day 20 and their sizes were determined. RESULTS: We observed that spheroids were formed within 24 hours regardless of the method and initial cell density. Further, in all groups the spheroid size was maximal at day 2, followed by a decline until day 20. Spheroids remained stable until day 20 independent of initial seeding concentration in all groups. DISCUSSION: We have generated long-term culture spheroids of SCC4 cells. The size of the spheroids can be influenced by varying the initial cell seeding density until day 20. This may be useful if different sizes of spheroids are required, e.g. in hypoxia research. Show more

Keywords: 3D culture, spheroids, SCC4 cell line, oral squamous cell carcinoma

DOI: 10.3233/JCB-179005

Citation: Journal of Cellular Biotechnology, vol. 3, no. 1, pp. 41-50, 2017

Authors: Wegmann, Marcel | Kalmbach, Sabrina | Schmidt, Jonas | Blessing, Frithjof | Wenzel, Folker

Article Type: Research Article

Abstract: INTRODUCTION: Besides a sodium chloride solution, a hydroxyethyl starch solution, and an immunoglobulin G solution, a human albumin solution is more frequently used in infusion solutions. To prevent negative influence of volume replacement solutions on hemostasis, additional administration of fibrinogen has been used in this assignment with the intention of keeping the plasmatic hemostasis stable. Therefore, in this study possible effects on hemostasis by plasma dilution with HES along with the effects of other clinically applied volume replacement solutions will be analyzed. MATERIAL AND METHODS: Whilst maintaining the concentration of fibrinogen the experiment used four different volume replacement …solutions in increments of 1 : 3, 1 : 6 and 1 : 11. To evaluate the changed hemostasis the coagulation parameters PT, PTT and thrombin time have been measured and assessed. RESULTS: A consistent increase of PT is observable in all four dilution solutions. The undiluted samples have a mean value of 11.44 sec. For example, the highest increase of PT of all dilution factors could be detected in samples treated with human albumin solution, where 19.54 sec was the mean value of the 1 : 3 dilution, whereas values of the 1 : 11 dilution nearly tripled (85.00 sec). Lower values could be achieved in measurements where solutions were diluted with NaCl and HES. However, PT values for the NaCl and HES dilutions (1 : 6) were 39.26 seconds and 36.55 seconds and have exceeded the corresponding human albumin dilution value. However, the steepest curves are achieved using immunoglobulin G solution as dilution agent. CONCLUSION: All results explicitly show that a dilution of the plasma results in worsening the hemostasis. However, the thrombin time can be considered as an exception, as even with a stronger dilution the solutions do not show an increase. Additionally, it could be established that the intrinsic factors are stronger influenced in their function than the extrinsic factors. Show more

Keywords: Coagulation parameters, plasma dilution, fibrinogen stabilization

DOI: 10.3233/JCB-179006

Citation: Journal of Cellular Biotechnology, vol. 3, no. 1, pp. 51-60, 2017

Authors: Martí, Yasmina | Nürnberg, Elina | Horschitz, Sandra | Hafner, Mathias | Schloss, Patrick | Meyer-Lindenberg, Andreas | Lau, Thorsten

Article Type: Research Article

Abstract: BACKGROUND: Major pathomechanisms underlying neurodegenerative diseases, such as Parkinson’s Disease, are still not well understood. Induced human pluripotent and rodent embryonic stem cells provide powerful disease models to address neurodegeneration-inducing pathomechanisms on a molecular and cellular level. OBJECTIVE: Our aim is to establish a refined protocol to generate healthy and patient donor stem cell-derived dopaminergic neurons to investigate neurodegenerative events in vitro . METHODS: Human healthy donor- and patient-derived induced pluripotent stem cells were differentiated into stable dopaminergic progenitor cell lines and further differentiated into dopaminergic neurons. Induced pluripotent stem cells, neuronal progenitors and terminally differentiated …neurons were characterized by confocal laser microscopy-based immunofluorescence analysis, live cell imaging demonstrating dopamine transporter-specific uptake of a fluorescent substrate and transcriptome analysis. RESULTS: Based on our immunofluorescence analysis, dopaminergic differentiation approaches predominantly yield dopaminergic neurons and GFAP-expressing glial cells. We detected a small partition of GABAergic neurons, yet neither serotonergic nor glutamatergic neurons. Dopaminergic neurons were successfully stained for pre- and postsynaptic and mitochondrial markers. Live cell imaging experiments verified dopamine transporter-dependent uptake of the fluorescent monoamine transporter substrate ASP+. CONCLUSION: Human stem cell-derived dopaminergic neurons are a suitable cellular system for fluorescence-based experimental approaches to address neurodegenerative events in vitro . Show more

Keywords: Induced pluripotent stem cells, neuronal differentiation, stem cell-derived neurons, dopamine transporter, mitochondria

DOI: 10.3233/JCB-179007

Citation: Journal of Cellular Biotechnology, vol. 3, no. 1, pp. 61-80, 2017