03 October 2018 : Animal Research
Primary Cultivation and Identification of Vascular Smooth Muscle Cells from the Spiral Modiolar Artery of Guinea Pigs
Jingjie Xiao12BCDF, Zhiping Zhang13BDE, Wei Zhang14BC, Lei Wu15CD, Liang Zhang12DE, Yang Wang12DF, Li Li21DG, Xinzhi Li16AEFG*, Ketao Ma2AEFGDOI: 10.12659/MSM.912606
Med Sci Monit 2018; 24: ANS7023-7034
Abstract
BACKGROUND: This article reports a method to obtain vascular smooth muscle cells (SMCs) from the spiral modiolar artery (SMA) of guinea pigs and provides materials for related experimental studies.
MATERIAL AND METHODS: SMA was separated from the cochlea of guinea pigs, digested with trypsin (1.25 g/L) and allowed to adhere in a 35-mm culture dish. The morphology of the sample was investigated, and the sample was identified by immunofluorescence analysis, flow cytometry, Western blot, and RT-PCR. Cell viability was calculated using trypan blue and flow cytometry. Whole-cell patch clamp was used to record the membrane input resistance (Rinput), reciprocal membrane input conductance (Ginput), membrane input capacitance (Cinput), and resting membrane potential (RP) of the SMCs.
RESULTS: Microscopy results showed that the cells had typical peak–valley growth pattern. The cell growth curve was similar to an S curve, and flow cytometry results showed that the cell apoptosis rate was less than 10%. Moreover, flow cytometry, immunofluorescent staining, Western blot and RT-PCR detected the specific and intensely positive expression of cell type-specific markers α-SM-actin, SM22α, calponin and desmin. Furthermore, following properties of the P3 and P6 cells were obtained: Rinput, 2611±356 and 2477±338 MΩ; Ginput, 0.454±0.071 and 0.273±0.037 ns; Cinput, 17.029±0.917 and 18.042±1.051 pF, and RP –20.602±1.503 and –22.192±1.905 mV.
CONCLUSIONS: Various highly purified SMCs were obtained from the SMA of guinea pigs. We provide an ideal experimental material for the study of the pathogenesis of diseases related to the circulation disturbances in the inner ear in vitro. The results can be used to evaluate the effects of drugs on vascular smooth muscle.
Keywords: Guinea Pigs, Myocytes, Smooth Muscle, primary cell culture
Editorial
01 June 2024 : Editorial
Editorial: Concerns as Highly Pathogenic Avian Influenza (HPAI) Virus of the H5N1 Subtype is Identified in Dairy Cows and Other MammalsDOI: 10.12659/MSM.945315
Med Sci Monit 2024; 30:e945315
In Press
Review article
A New Pandemic of the XXIst Century: The Growing Crisis of Adolescent Depression in the Digital AgeMed Sci Monit In Press; DOI: 10.12659/MSM.944838
Review article
A Review of Wastewater-Based Epidemiology Studies for the Assessment of Over-the-Counter Medicines Used as ...Med Sci Monit In Press; DOI: 10.12659/MSM.944120
Clinical Research
Functional Outcomes of Therapeutic Selective Nerve Root Block for Single-Segment Lumbar Spinal Stenosis: A ...Med Sci Monit In Press; DOI: 10.12659/MSM.943634
Clinical Research
Shortening Oral Simethicone-to-Colonoscopy Interval Increases Bowel Preparation QualityMed Sci Monit In Press; DOI: 10.12659/MSM.943972
Most Viewed Current Articles
17 Jan 2024 : Review article 1,724,770
Vaccination Guidelines for Pregnant Women: Addressing COVID-19 and the Omicron VariantDOI :10.12659/MSM.942799
Med Sci Monit 2024; 30:e942799
14 Dec 2022 : Clinical Research 1,528,873
Prevalence and Variability of Allergen-Specific Immunoglobulin E in Patients with Elevated Tryptase LevelsDOI :10.12659/MSM.937990
Med Sci Monit 2022; 28:e937990
16 May 2023 : Clinical Research 690,279
Electrophysiological Testing for an Auditory Processing Disorder and Reading Performance in 54 School Stude...DOI :10.12659/MSM.940387
Med Sci Monit 2023; 29:e940387
01 Jan 2022 : Editorial 50,227
Editorial: Current Status of Oral Antiviral Drug Treatments for SARS-CoV-2 Infection in Non-Hospitalized Pa...DOI :10.12659/MSM.935952
Med Sci Monit 2022; 28:e935952