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1. Human dermal fibroblast proliferation controlled by surface roughness of two-component nanostructured latex polymer coatings.

Author

Rosqvist E, Niemelä E, Venu AP, Kummala R, Ihalainen P, Toivakka M, Eriksson JE, and Peltonen J

Subjects
Cells, Cultured, Humans, Surface Properties, Cell Proliferation, Dermis cytology, Fibroblasts cytology, Latex chemistry, Nanostructures chemistry, Polymers chemistry
Abstract

In this study hierarchically-structured latex polymer coatings and self-supporting films were characterised and their suitability for cell growth studies was tested with Human Dermal Fibroblasts (HDF). Latex can be coated or printed on rigid or flexible substrates thus enabling high-throughput fabrication. Here, coverslip glass substrates were coated with blends of two different aqueous latex dispersions: hydrophobic polystyrene (PS) and hydrophilic carboxylated acrylonitrile butadiene styrene (ABS). The nanostructured morphology and topography of the latex films was controlled by varying the mixing ratio of the components in the latex blend. Thin latex-coatings retain high transparency on glass allowing optical and high resolution imaging of cell growth and morphology. Compared to coverslip glass surfaces and commercial well-plates HDF cell growth was enhanced up to 150-250 % on latex surfaces with specific nanostructure. Growth rates were correlated with selected roughness parameters such as effective surface area (S q ), RMS-roughness (S dr ) and correlation length (S cl37 ). High-resolution confocal microscopy clearly indicated less actin stress-fibre development in cells on the latex surface compared to coverslip glass. The results show that surface nanotopography can, by itself, passively modulate HDF cell proliferation and cytoskeletal architecture., (Copyright © 2018. Published by Elsevier B.V.)

Published
2019
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2. RNA Extraction from Ears and Draining Lymph Nodes of Mice Infected with Leishmania amazonensis

Author

Emilie Giraud, Evie Melanitou, Interactions Virus-Insectes - Insect-Virus Interactions (IVI), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Département Parasites et Insectes vecteurs - Department of Parasites and Insect Vectors, Institut Pasteur [Paris], This work was supported by institutional funding from the Institut Pasteur., We are grateful to the members of the Immunophysiology and Parasitism Unit for their recommendations for these experimental procedures and their participation to the original work described in the published paper from where this protocol is derived (Giraud et al. al., 2019b, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)

Subjects
Mouse, Strategy and Management, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Industrial and Manufacturing Engineering, Microbiology, Dermis, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, medicine, Methods Article, Parasite hosting, Vector (molecular biology), Ear pinna dermis, Lymph nodes, biology, Mechanical Engineering, Metals and Alloys, RNA, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, biology.organism_classification, medicine.disease, 3. Good health, Sandfly, RNA isolation, [SDV.GEN.GA]Life Sciences [q-bio]/Genetics/Animal genetics, medicine.anatomical_structure, Visceral leishmaniasis, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, RNA extraction, Lymph, Leishmania amazonensis
Abstract

International audience; Parasites of the genus Leishmania infect the mammalian hosts, including mice and humans and cause cutaneous or visceral leishm aniasis depending upon the parasite species transmitted by the vector sandfly. Leishmania amazonensis is one of the Leishmania species responsible for th e cutaneous form of the disease. We have inoculated with these parasites the ear dermis of mice. RNA preparations were performed from fragmented tissues using a buffer containing guanidin isothiocynate (RLT buffer, RNeasy Mini Kit, Qiagen, SAS, France) and β mercaptoethanol. Both reagents facilitate the isolation of intact RNA from tissues and the use of the RNeasy Kits present with several advantages that facilitate the isolation of pure non degraded total RNA: i) T his method allows to avoid the presence o f phenol in the RNA extraction buffer, commonly used in alternative protocols; ii) Moreover Diethylpyrocarbonate (DEPC) treatment of glassware, to avoid RNAses contamination of the samples, is not required with this protocol; iii) F inally, it is a fast pro cedure and the isolated total RNA may be concentrated in a small volume thus facilitating its use for downstream experimental procedures.

Published
2020
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