Mouse microglial cells (BV2), human microglia cells (HMC3), and mouse macrophage cells (RAW264.7) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China) and cultured in DMEM with 10% FBS (Thermo, Gibco, 10270106, United States). African green monkey kidney cells (Vero) were purchased from American Type Culture Collection (ATCC) and cultured in DMEM with 10% FBS (Hangzhou Sijiqing Biology Engineering Materials Co., Ltd., Hangzhou, China). All the cells were cultured at 37°C in a humid atmosphere with 5% CO2. HSV-1 F strain (ATCC, United States) is a gift from the University of Hong Kong and is stored at –80°C.
Hsv 1 strain f
Manufactured by American Type Culture Collection
Sourced in United States
HSV-1 strain F is a laboratory strain of the Herpes Simplex Virus Type 1. It serves as a reference material for research and diagnostic purposes.
Automatically generated - may contain errors
Lab products found in correlation
Vero cell, by American Type Culture Collection (5 mentions)
Fnd500, by ExCell Bio (5 mentions)
Sh sy5y, by American Type Culture Collection (3 mentions)
Hacat, by American Type Culture Collection (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Vero cell line, by American Type Culture Collection (2 mentions)
Raw264, by National Collection of Authenticated Cell Cultures (1 mentions)
U87mg, by Thermo Fisher Scientific (1 mentions)
Anti gapdh 2118, by Cell Signaling Technology (1 mentions)
Ma 104, by Thermo Fisher Scientific (1 mentions)
Sh sy5y, by Thermo Fisher Scientific (1 mentions)
Anti gb ab6506, by Abcam (1 mentions)
U87mg, by American Type Culture Collection (1 mentions)
Ma104, by American Type Culture Collection (1 mentions)
Sh sy5y cell, by American Type Culture Collection (1 mentions)
Sk n sh, by American Type Culture Collection (1 mentions)
Mrc 5 cells, by American Type Culture Collection (1 mentions)
Htb 11, by American Type Culture Collection (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
10 protocols using hsv 1 strain f
1
Cell Culture and Virus Maintenance
2
Oncolytic Herpes Simplex Virus Engineering
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The HSV-1 F strain and Vero cells were purchased from ATCC. The oncolytic VAE viruses (γ34.5−, ICP6−, endo–angio+) and r-HSV-1 (γ34.5−, ICP6−; used as a control virus) were constructed by Dr. Willam Jia. The structures of the virus have been previously described [26] (link). The r-HSV-1 genome consists of long and short unique regions (UL and US), each bounded by terminal (T) and internal (I) repeat regions (RL and RS). The virus was engineered from the wild-type HSV-1 strain F by deleting 1 kb within both copies of the γ34.5 gene. VAE, an oncolytic r-HSV virus, was derived from r-HSV by inserting the endo–angio fusion gene into the ICP6 coding region. This virus was produced in Vero cellsas previously described [28] (link). The final experimental grade virus preparation was titrated for infectivity, tested for contamination, and stored at −70°C in identical pairs of vials. Each vial contained the virus at the appropriate concentration (107 plaque-forming units, p.f.u.) in a 1 ml total volume of human serum albumin (HSA, 0.5% to 1.0%) in isotonic PBS. The virus was titrated using standard methods in BHK21/C13 cells. Viral titers were expressed as the concentration (p.f.u.) compared with the TCID50 values.
3
Cultivation of Cell Lines and Viruses
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Kidney epithelial cells MA-104 (CRL-2378.1) and Vero (CCL-81), neuronal cells SH-SY5Y (CRL-2266) and U87-MG (HTB-14) were obtained from the American Type Culture Collection Center (ATCC, USA). U87-MG, Vero, and SH-SY5Y cells were cultured in Dulbecco's modified Eagle's medium (8118305, GIBCO, USA), supplemented with 10% fetal bovine serum (FND500, ExCell Bio, Shanghai, China). MA-104 cells were cultured in modified Eagle's medium (A1451801, GIBCO, USA).
HSV-1 F strain (ATCC, USA) was grown in Vero cells and stored at − 80 °C for further use as previously described [16 (link)]. ACV-resistant strains HSV-1/153 and HSV-1/Blue were obtained from the Guangzhou Institutes of Biomedicine and Health (Guangzhou, China). HSV-1 F strain tagged by Green fluorescent protein (GFP) (GFP-HSV-1) was obtained from Professor Yuan Li of Jinan University (Guangzhou, China). HSV-2 and RV virus were obtained from the Wuhan Institute of Virology (Wuhan, China). As previously described, the influenza virus H1N1 was grown and stored at − 80 °C for further use [17 (link)]. Antibody anti-ICP0 (ab6513) and anti-gB (ab6506) were purchased from Abcam (Cambridge, UK), and anti-GAPDH (2118) was purchased from Cell Signaling Technology (Danvers, MA, USA).
HSV-1 F strain (ATCC, USA) was grown in Vero cells and stored at − 80 °C for further use as previously described [16 (link)]. ACV-resistant strains HSV-1/153 and HSV-1/Blue were obtained from the Guangzhou Institutes of Biomedicine and Health (Guangzhou, China). HSV-1 F strain tagged by Green fluorescent protein (GFP) (GFP-HSV-1) was obtained from Professor Yuan Li of Jinan University (Guangzhou, China). HSV-2 and RV virus were obtained from the Wuhan Institute of Virology (Wuhan, China). As previously described, the influenza virus H1N1 was grown and stored at − 80 °C for further use [17 (link)]. Antibody anti-ICP0 (ab6513) and anti-gB (ab6506) were purchased from Abcam (Cambridge, UK), and anti-GAPDH (2118) was purchased from Cell Signaling Technology (Danvers, MA, USA).
4
Established Cell Lines and Virus Stocks
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African green monkey kidney cell line (Vero cells) and Human immortalized keratinocyte cell line (HaCaT) were purchased from the American Type Culture Collection Center (ATCC). Vero, HaCaT, SH-SY5Y (ATCC CRL-2266), and BV2 (Cell Bank, Chinese Academy of Sciences) cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; 8118305, GIBCO/Thermo Fisher Scientific, United States) with 10% fetal bovine serum (FND500, ExCell Bio, Shanghai, China). HSV-1 F strain (ATCC, United States) preserved by the University of Hong Kong, was propagated in Vero cells and stored in the refrigerator at −80°C. HSV-1/Blue, a TK mutant derived from HSV-1 KOS strain and two clinical ACV-resistant HSV-1 strain, HSV-1/106 and HSV-1/153, were kind gifts from Prof Tao Peng (Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, China). Green fluorescent protein (GFP)-tagged HSV-1 F strain (GFP-HSV-1) (GFP-tagged viral protein Us11) was obtained from the research group of Professor Yuan Li (Jinan University, China).
5
Cell Culture Protocols for Viral Studies
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African green monkey kidney cell line (Vero cells) and Human immortalized keratinocyte cell line (HaCaT) were purchased from the American Type Culture Collection Center (ATCC) and cultured in Dulbecco’s Modified Eagle Medium (DMEM; 8118305, GIBCO/Thermo Fisher Scientific, Waltham, MA, USA) with 10% fetal bovine serum (FND500, Excell Bio, Shanghai, China). SH-SY5Y cells (ATCC CRL-2266) were cultured at 37 °C in a humid atmosphere with 5% CO2. HSV-1/Blue, a TK mutant derived from HSV-1 (KOS), and two acyclovir-resistant clinical HSV-1 strains, HSV-1/106 and HSV-1/153, were kind gifts from Tao Peng [20 (link)], State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences. GFP-HSV-1, expressing a GFP-tagged viral protein Us11, was used to evaluate viral-nuclear transport; HSV-1 strain F (ATCC, Manassas, VA, USA), initially obtained from Hong Kong University, was propagated in Vero cells and stored at −80 °C.
6
Investigating HSV-1 Dynamics in Neuroblastoma
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Vero cell line (ATCC, USA) was cultured in Dulbecco’s modified Eagle’s medium (DMEM; 8118305, GIBCO/Thermo Fisher Scientific, USA) with 10% fetal bovine serum (FND500, ExCell Bio, Shanghai, China). The neuroblastoma cell line SK-N-SH (ATCC, HTB-11, American Type Culture Collection, Manassas, VA, USA) was propagated in Eagle’s minimal essential medium (MEM; GIBCO /Thermo Fisher Scientific, USA) supplemented with 10% FBS. In our previous studies, HSV-1 infection induced the biphasic F-actin dynamics in SK-N-SH cells [26 (link)]. In this study, the SK-N-SH cells were used to study the effect of amentoflavone on F-actin. HSV-1 strain F (ATCC, USA), initially obtained from Hong Kong University, was propagated in Vero cells and stored at −80 °C until use. HSV-1/Blue, a TK mutant derived from HSV-1 (KOS) and two acyclovir-resistant clinical HSV-1 strains HSV-1/106 and HSV-1/153 were kind gifts from Tao Peng, State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences. GFP-HSV-1, expressing a GFP-tagged viral protein Us11, was used to evaluate viral nuclear transport [27 (link)]. HSV-1 Us11 is a multifunctional late protein which can regulate the accumulation of RNA species and facilitate HSV-1 replication [28 (link)].
7
Cultivating Cells for HSV-1 Studies
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MRC-5 cells (ATCC) and Vero cells (ATCC) were cultured as described previously [15] (link). All experiments were performed with the HSV-1 strain F (ATCC), a kind gift from Hong Kong University. The clinical-isolated ACV-resistant HSV-1 strain (named C106) used in this work was obtained from the Guangzhou Institutes of Biomedicine and Health [16] (link).
8
Vero and SH-SY5Y Cell Culture for HSV-1
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Vero cell line (ATCC, Manassas, VA, USA) was cultured in Dulbecco’s modified Eagle’s medium (DMEM; 8118305, GIBCO/Thermo Fisher Scientific, Gaithersburg, MD, USA) with 10% fetal bovine serum (11011-8611, TIANHANG, Hangzhou, China). SH-SY5Y (ATCC, Manassas, VA, USA), a human neuroblastoma cell line was cultured in Dulbecco’s modified Eagle’s medium (DMEM; 8118305, GIBCO/Thermo Fisher Scientific, Gaithersburg, MD, USA) with 10% fetal bovine serum (FND500, ExCell Bio, Shanghai, China). HSV-1 strain F (ATCC, Manassas, VA, USA), initially obtained from Hong Kong University, was propagated in Vero cells and stored at −80 °C until use. EGFP-HSV-1, expressing an EGFP-tagged viral protein Us11, a kind gift from the College of Pharmacy, Jinan University (Guangzhou, China), was used to evaluate viral replication capacity [18 (link)].
9
Propagation of Neuroblastoma and HSV-1
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The sources and culture conditions for different cell lines have been previously described (7 (link), 37 (link)). The neuroblastoma cell line SK–N–SH (ATCC HTB-11), purchased from the cell bank of the Chinese Academy of Sciences, was propagated in Eagle’s minimal essential medium (MEM) (Invitrogen) supplemented with 10% fetal bovine serum (FBS) (Invitrogen), 1.0 mM sodium pyruvate (Sigma), 0.1 mM nonessential amino acids (Invitrogen), and 1.5 g/liter sodium bicarbonate (Sigma). HSV-1 strain F (ATCC VR733), obtained from Hong Kong University, was propagated in Vero cells and stored at −80°C until use. All cells were purchased from ATCC.
10
Myricetin Antiviral Activity Evaluation
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Myricetin (with purity > 95%) was purchased from Topscience Co., Ltd. (Shanghai, China). Acyclovir was purchased from Sigma Aldrich (St. Louis, MO, USA). Vero, HeLa and Hep-2 cells were routinely cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (ExCell Bio, China), penicillin (100 U/mL), and streptomycin (100 μg/mL) at 37 °C in 5% CO2. HSV-1 strain F was purchased from ATCC (VR-734). HSV-2 strain 333 was obtained from Wuhan Institute of Virology, Chinese Academy of Sciences.
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