Endothelial Cells of the Lung
Joan Gil in Models of Lung Disease, 2020
There is obvious value in being able to isolate and maintain differentiated lung endothelial cells in culture that can be used for in vitro studies. While cell culture has some obvious advantages over whole animal studies, one must always be concerned about the artifactual nature of cells maintained in vitro. In most instances, cells are grown on a plastic substrate that is distinctly different from what they normally experience in vivo. A number of approaches can be used to mimic more faithfully the extracellular environment that cells normally interact with: (1) maintaining cells on purified matrix proteins that have been coated on the cell culture dish, (2) using a three-dimensional “native” collagen matrix, and (3) growing endothelial cells on a matrix previously deposited by endothelial cells. With the use of such approaches it has been demonstrated, in a variety of cell systems, that the extracellular matrix environment of cells plays an important role in defining their phenotypic expression (Macarak and Howard, 1983; Canfield et al., 1986; Grinnell, 1982; Bissell et al., 1982).
Alternative Methodologies to Animal Testing
Nicola Loprieno in Alternative Methodologies for the Safety Evaluation of Chemicals in the Cosmetic Industry, 2019
Examples of possible tests are represented by: Cell Cultures cell linesprimary cell cultureOrgan Cultures nonmammalianmammalianWhole Embryo Cultures
The Thymus in the Regulation and Control of Cell Growth
Nate F. Cardarelli in The Thymus in Health and Senescence, 2019
Le Douarin and Jotereau, in discussing the homing of lymphoid stem cells of the quail make note of the fact that hemopoietic cells colonize in waves.548 Movement of such cells to the thymus and bursa are periodic, implying some sort of endogenous control or programming. There are other indications of thymus involved regulation in embryonic life. Some I have previously described. The shifting the types of cortical thymocyte specific antigens with fetal age, for instance.549,550 Changes in thymic gangliosides have been observed, and these substances may serve as a marker for immature thymus cells.551 The neonatal thymic cell repertoire varies considerably from that of the mature thymus. Certain suppressor cells are missing, for instance.552In vitro cell culture indicates considerable morphological and biochemical distinctions between cell types.553 In the mouse embryo, dramatic cell changes are observed at about the 17th or 18th day of fetal life, sympathetic innervation occurring about day 17.554 Consequently, such changes likely require nervous stimulation. Various neurotransmitters are known to promote mitosis in cultured thymic lymphocytes.555
Chrysin Suppresses HT-29 Cell Death Induced by Diclofenac through Apoptosis and Oxidative Damage
Published in Nutrition and Cancer, 2021
The assay was performed following the procedure supplied with the ApopTag kit (Milipore). The dye used in the assay specifically target 3′-OH DNA breaks in apoptotic cells. Terminal deoxynucleotidyl transferase catalyzes the addition of deoxyribonucleotide triphosphate to the 3′-OH end of DNA breaks stemmed from apoptosis. This distinguish apoptosis from necrosis by specifically detecting DNA cleavage and chromatin condensation associated with apoptosis. The HT-29 were seeded in a 6-well plate covering approximately 5 × 105 cells/well. The cells were grown as described in cell culture and treatment. Later, the HT-29 were observed under an inverted light microscope. At least three pictures were obtained for each treated group and Apoptotic Index (AI) was calculated as described previously (20).
Evaluation of Calu-3 cell lines as an in vitro model to study the inhalation toxicity of flavoring extracts
Published in Toxicology Mechanisms and Methods, 2022
Xiaoli Ji, Yunhua Sheng, Ying Guan, Yinxia Li, Yuqiong Xu, Liming Tang
In summary, several in vitro models have been developed to study the potential toxicity of inhaled materials, and only a few studies have been performed on the Calu-3 cell line, which is representative of the bronchial epithelial barrier. Much remains to be elucidated concerning the function, structure, and toxicity responses of airway epithelia. This study suggests that culturing Calu-3 cells under ALI conditions and culture times of 8–10 days, cell morphology, and barrier function were optimized in an in vitro model. Under these conditions, cells can be successfully cultured with good cell culture growth and differentiation. Therefore, the Calu-3 cell line has considerable potential as an in vitro model for bronchial epithelium. In vitro evidence concerning the respiratory toxicity caused by e-cigarette-induced flavoring chemicals is scarce, and a reliable in vitro model is required to evaluate their inhalation toxicity.
Targeting mitochondria in dermatological therapy: beyond oxidative damage and skin aging
Published in Expert Opinion on Therapeutic Targets, 2022
Tongyu C Wikramanayake, Jérémy Chéret, Alec Sevilla, Mark Birch-Machin, Ralf Paus
It is perfectly reasonable to expect that these well-known regulators of mitochondrial activity and/or biogenesis also operate in human skin and that standard dermatotherapeutics like glucosteroids, retinoids and vitamin D3 derivates all impact profoundly on these (Table 2). Yet, how exactly mitochondrial function is regulated in human skin remains very incompletely understood, namely in proliferating/differentiating epidermal and HF keratinocytes, as well as in sebocytes in vivo. Cell culture studies provide some pointers. For example, the vitamin D receptor is imported into the mitochondria in a ligand-independent manner in cultured HaCaT human keratinocytes [282], colocalizes with StAR (STARD1) and Cytochrome P450 Family 11 Subfamily A Member 1 (CYP11A1) within the mitochondria and shows inhibition of mitochondrial respiration and ATP production [1,25(OH)2D3 > 20(OH)D3] [283]. In differentiating keratinocytes, vitamin D treatment promotes intracellular lipid deposition, but in proliferating HaCaT, this biosynthetic pathway was not inducible by the hormone [284]. However, it is unclear to what extent these in vitro findings translate to human skin physiology.
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