cell culture ectocervical Search Results


ectocervical  (ATCC)
96
ATCC ectocervical
Ectocervical, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ectocervical primary cell medium  (Thermo Fisher)
99
Thermo Fisher ectocervical primary cell medium
a Schematic representation of human <t>ectocervical</t> organoid and 2D stem cell culture generation from biopsies. b – d Confocal images of human ectocervical tissue and organoids immunolabeled for KRT5, KRT8 (b); Ki67, E-cadherin (CDH1) ( c ); Ki67, p63 ( d ); Nuclei in blue. e PCR to detect HPV16, HPV18 E6E7, and GAPDH in samples from three independent experiments. Donor 1–3, untreated ectocervical stem cells; mock, Polybrene treated cells; Donor 1-3 HPV E6E7, Donor 1-3 transduced with HPV16 E6E7 lentivirus; End1, End1 cells; HeLa, HeLa cells; Donor 3 Luci, cells transduced with Luciferase lentivirus; J23T3, feeder cells; water, water control. f , g Confocal images of ectocervical organoids expressing HPV16 E6E7 immunolabeled for KRT5, KRT8 ( f ); Ki67, CDH1 ( g ); Nuclei are shown in blue. Images in ( b – d ) and ( f , g ) represent organoid cultures from three biological experiments. h Lysates of hCEcto and hCEcto E6E7 organoids were subjected to immunoblot analysis for CDH1 and β-actin as a loading control. Data represent three biological replicates. Densitometry values for CDH1 normalized to the β-actin, values represent the relative FC in hCEcto E6E7 compared to hCEcto. i , j Proliferation and stemness were measured by quantification of the diameter of hCEcto and hCEcto E6E7 organoids ( i ) and organoid formation ( j ). Data are presented as violin plot min–max, lines are 25 and 75th percentile, bold line is median ( i ); data are presented as mean values ± SD of four biological replicates ( j ). k Relative mRNA expression of HPV16 E6E7 analyzed by qRT-PCR from RNA extracts of three Donors (1–3) HPV E6E7, W12-Int or W12-Epi, and KC-Epi cells. Whiskers are min–max, box is 25–75th percentile, lines are median values normalized to the geometric mean of Donor 1–3 HPV E6E7. l – n smRNA-ISH images of organoids derived from hCEcto ( l ), hCEcto E6E7 ( m ), and KC-Epi ( n ) probed with HPV16 L1 and E7; Nuclei are shown in blue. Images are representatives of three replicates. Statistical significance was calculated using ( i , j ) two-sided t test, ( k ) One-way ANOVA, P -values are indicated. Source data are provided as a Source Data file.
Ectocervical Primary Cell Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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endocervical epithelium derived cell lines vk2  (ATCC)
96
ATCC endocervical epithelium derived cell lines vk2
Cytotoxicity and antiviral activity of Avirulins in female reproductive tract <t>epithelium.</t> ( a – c ) Immortalized epithelial cell lines derived from the vaginal wall, ectocervix, and endocervix <t>(VK2,</t> Ect1, and End1, respectively) were treated with Avirulin lead compounds Av-5, Av-14, and Av-26 at a range of concentrations from 0.78–100 µM for 24 h. Cytotoxicity was measured using a commercially available luminescence-based cytotoxicity assay, CytotoxGlo. Cytotoxicity of treatments was compared to DMSO vehicle control, with the detergent Triton-X 100 for complete cell death. ( n = 4, error = SEM). ( d ) Av-5 was incubated with clarified vaginal fluid at 37 °C for 2 h, then diluted with culture media and applied to TZM-bl. Cells were infected with BaL for 24 h, then inhibition was determined using the luciferase reporter assay. Cervico–vaginal fluid from healthy women is inherently antiviral, as shown in the vehicle control. n = 3, error = SEM * = p > 0.05. ( e ) Cytotoxicity of vaginal fluids and 5 µM Av-5 on TZM-bl measured using CytotoxGlo assay. Cervico–vaginal fluid (CVF) dilutions with Av-5 or equivalent DMSO were incubated at 37 °C for 2 h, then applied to TZM-bl. Cell death at 24 h was determined by subtraction of total cell luminescence from dead cell luminescence. n = 3, error = SEM.
Endocervical Epithelium Derived Cell Lines Vk2, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ectocervical epithelial cell line  (ATCC)
97
ATCC ectocervical epithelial cell line
Cytotoxicity and antiviral activity of Avirulins in female reproductive tract <t>epithelium.</t> ( a – c ) Immortalized epithelial cell lines derived from the vaginal wall, ectocervix, and endocervix <t>(VK2,</t> Ect1, and End1, respectively) were treated with Avirulin lead compounds Av-5, Av-14, and Av-26 at a range of concentrations from 0.78–100 µM for 24 h. Cytotoxicity was measured using a commercially available luminescence-based cytotoxicity assay, CytotoxGlo. Cytotoxicity of treatments was compared to DMSO vehicle control, with the detergent Triton-X 100 for complete cell death. ( n = 4, error = SEM). ( d ) Av-5 was incubated with clarified vaginal fluid at 37 °C for 2 h, then diluted with culture media and applied to TZM-bl. Cells were infected with BaL for 24 h, then inhibition was determined using the luciferase reporter assay. Cervico–vaginal fluid from healthy women is inherently antiviral, as shown in the vehicle control. n = 3, error = SEM * = p > 0.05. ( e ) Cytotoxicity of vaginal fluids and 5 µM Av-5 on TZM-bl measured using CytotoxGlo assay. Cervico–vaginal fluid (CVF) dilutions with Av-5 or equivalent DMSO were incubated at 37 °C for 2 h, then applied to TZM-bl. Cell death at 24 h was determined by subtraction of total cell luminescence from dead cell luminescence. n = 3, error = SEM.
Ectocervical Epithelial Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ectocervical epithelial cells ca ski cells  (ATCC)
97
ATCC ectocervical epithelial cells ca ski cells
Cytotoxicity and antiviral activity of Avirulins in female reproductive tract <t>epithelium.</t> ( a – c ) Immortalized epithelial cell lines derived from the vaginal wall, ectocervix, and endocervix <t>(VK2,</t> Ect1, and End1, respectively) were treated with Avirulin lead compounds Av-5, Av-14, and Av-26 at a range of concentrations from 0.78–100 µM for 24 h. Cytotoxicity was measured using a commercially available luminescence-based cytotoxicity assay, CytotoxGlo. Cytotoxicity of treatments was compared to DMSO vehicle control, with the detergent Triton-X 100 for complete cell death. ( n = 4, error = SEM). ( d ) Av-5 was incubated with clarified vaginal fluid at 37 °C for 2 h, then diluted with culture media and applied to TZM-bl. Cells were infected with BaL for 24 h, then inhibition was determined using the luciferase reporter assay. Cervico–vaginal fluid from healthy women is inherently antiviral, as shown in the vehicle control. n = 3, error = SEM * = p > 0.05. ( e ) Cytotoxicity of vaginal fluids and 5 µM Av-5 on TZM-bl measured using CytotoxGlo assay. Cervico–vaginal fluid (CVF) dilutions with Av-5 or equivalent DMSO were incubated at 37 °C for 2 h, then applied to TZM-bl. Cell death at 24 h was determined by subtraction of total cell luminescence from dead cell luminescence. n = 3, error = SEM.
Ectocervical Epithelial Cells Ca Ski Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ectocervical ect e6e7 epithelial cells lines  (ATCC)
95
ATCC ectocervical ect e6e7 epithelial cells lines
Cytotoxicity and antiviral activity of Avirulins in female reproductive tract <t>epithelium.</t> ( a – c ) Immortalized epithelial cell lines derived from the vaginal wall, ectocervix, and endocervix <t>(VK2,</t> Ect1, and End1, respectively) were treated with Avirulin lead compounds Av-5, Av-14, and Av-26 at a range of concentrations from 0.78–100 µM for 24 h. Cytotoxicity was measured using a commercially available luminescence-based cytotoxicity assay, CytotoxGlo. Cytotoxicity of treatments was compared to DMSO vehicle control, with the detergent Triton-X 100 for complete cell death. ( n = 4, error = SEM). ( d ) Av-5 was incubated with clarified vaginal fluid at 37 °C for 2 h, then diluted with culture media and applied to TZM-bl. Cells were infected with BaL for 24 h, then inhibition was determined using the luciferase reporter assay. Cervico–vaginal fluid from healthy women is inherently antiviral, as shown in the vehicle control. n = 3, error = SEM * = p > 0.05. ( e ) Cytotoxicity of vaginal fluids and 5 µM Av-5 on TZM-bl measured using CytotoxGlo assay. Cervico–vaginal fluid (CVF) dilutions with Av-5 or equivalent DMSO were incubated at 37 °C for 2 h, then applied to TZM-bl. Cell death at 24 h was determined by subtraction of total cell luminescence from dead cell luminescence. n = 3, error = SEM.
Ectocervical Ect E6e7 Epithelial Cells Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ectocervical cells  (ATCC)
99
ATCC ectocervical cells
Cytotoxicity and antiviral activity of Avirulins in female reproductive tract <t>epithelium.</t> ( a – c ) Immortalized epithelial cell lines derived from the vaginal wall, ectocervix, and endocervix <t>(VK2,</t> Ect1, and End1, respectively) were treated with Avirulin lead compounds Av-5, Av-14, and Av-26 at a range of concentrations from 0.78–100 µM for 24 h. Cytotoxicity was measured using a commercially available luminescence-based cytotoxicity assay, CytotoxGlo. Cytotoxicity of treatments was compared to DMSO vehicle control, with the detergent Triton-X 100 for complete cell death. ( n = 4, error = SEM). ( d ) Av-5 was incubated with clarified vaginal fluid at 37 °C for 2 h, then diluted with culture media and applied to TZM-bl. Cells were infected with BaL for 24 h, then inhibition was determined using the luciferase reporter assay. Cervico–vaginal fluid from healthy women is inherently antiviral, as shown in the vehicle control. n = 3, error = SEM * = p > 0.05. ( e ) Cytotoxicity of vaginal fluids and 5 µM Av-5 on TZM-bl measured using CytotoxGlo assay. Cervico–vaginal fluid (CVF) dilutions with Av-5 or equivalent DMSO were incubated at 37 °C for 2 h, then applied to TZM-bl. Cell death at 24 h was determined by subtraction of total cell luminescence from dead cell luminescence. n = 3, error = SEM.
Ectocervical Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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normal ectocervical hec cell lines  (ATCC)
97
ATCC normal ectocervical hec cell lines
Cytotoxicity and antiviral activity of Avirulins in female reproductive tract <t>epithelium.</t> ( a – c ) Immortalized epithelial cell lines derived from the vaginal wall, ectocervix, and endocervix <t>(VK2,</t> Ect1, and End1, respectively) were treated with Avirulin lead compounds Av-5, Av-14, and Av-26 at a range of concentrations from 0.78–100 µM for 24 h. Cytotoxicity was measured using a commercially available luminescence-based cytotoxicity assay, CytotoxGlo. Cytotoxicity of treatments was compared to DMSO vehicle control, with the detergent Triton-X 100 for complete cell death. ( n = 4, error = SEM). ( d ) Av-5 was incubated with clarified vaginal fluid at 37 °C for 2 h, then diluted with culture media and applied to TZM-bl. Cells were infected with BaL for 24 h, then inhibition was determined using the luciferase reporter assay. Cervico–vaginal fluid from healthy women is inherently antiviral, as shown in the vehicle control. n = 3, error = SEM * = p > 0.05. ( e ) Cytotoxicity of vaginal fluids and 5 µM Av-5 on TZM-bl measured using CytotoxGlo assay. Cervico–vaginal fluid (CVF) dilutions with Av-5 or equivalent DMSO were incubated at 37 °C for 2 h, then applied to TZM-bl. Cell death at 24 h was determined by subtraction of total cell luminescence from dead cell luminescence. n = 3, error = SEM.
Normal Ectocervical Hec Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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normal ectocervical hec cell lines - by Bioz Stars, 2026-04
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epithelial ectocervical cell line  (ATCC)
93
ATCC epithelial ectocervical cell line
Cytotoxicity and antiviral activity of Avirulins in female reproductive tract <t>epithelium.</t> ( a – c ) Immortalized epithelial cell lines derived from the vaginal wall, ectocervix, and endocervix <t>(VK2,</t> Ect1, and End1, respectively) were treated with Avirulin lead compounds Av-5, Av-14, and Av-26 at a range of concentrations from 0.78–100 µM for 24 h. Cytotoxicity was measured using a commercially available luminescence-based cytotoxicity assay, CytotoxGlo. Cytotoxicity of treatments was compared to DMSO vehicle control, with the detergent Triton-X 100 for complete cell death. ( n = 4, error = SEM). ( d ) Av-5 was incubated with clarified vaginal fluid at 37 °C for 2 h, then diluted with culture media and applied to TZM-bl. Cells were infected with BaL for 24 h, then inhibition was determined using the luciferase reporter assay. Cervico–vaginal fluid from healthy women is inherently antiviral, as shown in the vehicle control. n = 3, error = SEM * = p > 0.05. ( e ) Cytotoxicity of vaginal fluids and 5 µM Av-5 on TZM-bl measured using CytotoxGlo assay. Cervico–vaginal fluid (CVF) dilutions with Av-5 or equivalent DMSO were incubated at 37 °C for 2 h, then applied to TZM-bl. Cell death at 24 h was determined by subtraction of total cell luminescence from dead cell luminescence. n = 3, error = SEM.
Epithelial Ectocervical Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ectocervical cell line ect1 culture t vaginalis strain ru393  (ATCC)
93
ATCC ectocervical cell line ect1 culture t vaginalis strain ru393
Cytotoxicity and antiviral activity of Avirulins in female reproductive tract <t>epithelium.</t> ( a – c ) Immortalized epithelial cell lines derived from the vaginal wall, ectocervix, and endocervix <t>(VK2,</t> Ect1, and End1, respectively) were treated with Avirulin lead compounds Av-5, Av-14, and Av-26 at a range of concentrations from 0.78–100 µM for 24 h. Cytotoxicity was measured using a commercially available luminescence-based cytotoxicity assay, CytotoxGlo. Cytotoxicity of treatments was compared to DMSO vehicle control, with the detergent Triton-X 100 for complete cell death. ( n = 4, error = SEM). ( d ) Av-5 was incubated with clarified vaginal fluid at 37 °C for 2 h, then diluted with culture media and applied to TZM-bl. Cells were infected with BaL for 24 h, then inhibition was determined using the luciferase reporter assay. Cervico–vaginal fluid from healthy women is inherently antiviral, as shown in the vehicle control. n = 3, error = SEM * = p > 0.05. ( e ) Cytotoxicity of vaginal fluids and 5 µM Av-5 on TZM-bl measured using CytotoxGlo assay. Cervico–vaginal fluid (CVF) dilutions with Av-5 or equivalent DMSO were incubated at 37 °C for 2 h, then applied to TZM-bl. Cell death at 24 h was determined by subtraction of total cell luminescence from dead cell luminescence. n = 3, error = SEM.
Ectocervical Cell Line Ect1 Culture T Vaginalis Strain Ru393, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ectocervical stem cells  (ATCC)
99
ATCC ectocervical stem cells
Cytotoxicity and antiviral activity of Avirulins in female reproductive tract <t>epithelium.</t> ( a – c ) Immortalized epithelial cell lines derived from the vaginal wall, ectocervix, and endocervix <t>(VK2,</t> Ect1, and End1, respectively) were treated with Avirulin lead compounds Av-5, Av-14, and Av-26 at a range of concentrations from 0.78–100 µM for 24 h. Cytotoxicity was measured using a commercially available luminescence-based cytotoxicity assay, CytotoxGlo. Cytotoxicity of treatments was compared to DMSO vehicle control, with the detergent Triton-X 100 for complete cell death. ( n = 4, error = SEM). ( d ) Av-5 was incubated with clarified vaginal fluid at 37 °C for 2 h, then diluted with culture media and applied to TZM-bl. Cells were infected with BaL for 24 h, then inhibition was determined using the luciferase reporter assay. Cervico–vaginal fluid from healthy women is inherently antiviral, as shown in the vehicle control. n = 3, error = SEM * = p > 0.05. ( e ) Cytotoxicity of vaginal fluids and 5 µM Av-5 on TZM-bl measured using CytotoxGlo assay. Cervico–vaginal fluid (CVF) dilutions with Av-5 or equivalent DMSO were incubated at 37 °C for 2 h, then applied to TZM-bl. Cell death at 24 h was determined by subtraction of total cell luminescence from dead cell luminescence. n = 3, error = SEM.
Ectocervical Stem Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


a Schematic representation of human ectocervical organoid and 2D stem cell culture generation from biopsies. b – d Confocal images of human ectocervical tissue and organoids immunolabeled for KRT5, KRT8 (b); Ki67, E-cadherin (CDH1) ( c ); Ki67, p63 ( d ); Nuclei in blue. e PCR to detect HPV16, HPV18 E6E7, and GAPDH in samples from three independent experiments. Donor 1–3, untreated ectocervical stem cells; mock, Polybrene treated cells; Donor 1-3 HPV E6E7, Donor 1-3 transduced with HPV16 E6E7 lentivirus; End1, End1 cells; HeLa, HeLa cells; Donor 3 Luci, cells transduced with Luciferase lentivirus; J23T3, feeder cells; water, water control. f , g Confocal images of ectocervical organoids expressing HPV16 E6E7 immunolabeled for KRT5, KRT8 ( f ); Ki67, CDH1 ( g ); Nuclei are shown in blue. Images in ( b – d ) and ( f , g ) represent organoid cultures from three biological experiments. h Lysates of hCEcto and hCEcto E6E7 organoids were subjected to immunoblot analysis for CDH1 and β-actin as a loading control. Data represent three biological replicates. Densitometry values for CDH1 normalized to the β-actin, values represent the relative FC in hCEcto E6E7 compared to hCEcto. i , j Proliferation and stemness were measured by quantification of the diameter of hCEcto and hCEcto E6E7 organoids ( i ) and organoid formation ( j ). Data are presented as violin plot min–max, lines are 25 and 75th percentile, bold line is median ( i ); data are presented as mean values ± SD of four biological replicates ( j ). k Relative mRNA expression of HPV16 E6E7 analyzed by qRT-PCR from RNA extracts of three Donors (1–3) HPV E6E7, W12-Int or W12-Epi, and KC-Epi cells. Whiskers are min–max, box is 25–75th percentile, lines are median values normalized to the geometric mean of Donor 1–3 HPV E6E7. l – n smRNA-ISH images of organoids derived from hCEcto ( l ), hCEcto E6E7 ( m ), and KC-Epi ( n ) probed with HPV16 L1 and E7; Nuclei are shown in blue. Images are representatives of three replicates. Statistical significance was calculated using ( i , j ) two-sided t test, ( k ) One-way ANOVA, P -values are indicated. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Modelling Chlamydia and HPV co-infection in patient-derived ectocervix organoids reveals distinct cellular reprogramming

doi: 10.1038/s41467-022-28569-1

Figure Lengend Snippet: a Schematic representation of human ectocervical organoid and 2D stem cell culture generation from biopsies. b – d Confocal images of human ectocervical tissue and organoids immunolabeled for KRT5, KRT8 (b); Ki67, E-cadherin (CDH1) ( c ); Ki67, p63 ( d ); Nuclei in blue. e PCR to detect HPV16, HPV18 E6E7, and GAPDH in samples from three independent experiments. Donor 1–3, untreated ectocervical stem cells; mock, Polybrene treated cells; Donor 1-3 HPV E6E7, Donor 1-3 transduced with HPV16 E6E7 lentivirus; End1, End1 cells; HeLa, HeLa cells; Donor 3 Luci, cells transduced with Luciferase lentivirus; J23T3, feeder cells; water, water control. f , g Confocal images of ectocervical organoids expressing HPV16 E6E7 immunolabeled for KRT5, KRT8 ( f ); Ki67, CDH1 ( g ); Nuclei are shown in blue. Images in ( b – d ) and ( f , g ) represent organoid cultures from three biological experiments. h Lysates of hCEcto and hCEcto E6E7 organoids were subjected to immunoblot analysis for CDH1 and β-actin as a loading control. Data represent three biological replicates. Densitometry values for CDH1 normalized to the β-actin, values represent the relative FC in hCEcto E6E7 compared to hCEcto. i , j Proliferation and stemness were measured by quantification of the diameter of hCEcto and hCEcto E6E7 organoids ( i ) and organoid formation ( j ). Data are presented as violin plot min–max, lines are 25 and 75th percentile, bold line is median ( i ); data are presented as mean values ± SD of four biological replicates ( j ). k Relative mRNA expression of HPV16 E6E7 analyzed by qRT-PCR from RNA extracts of three Donors (1–3) HPV E6E7, W12-Int or W12-Epi, and KC-Epi cells. Whiskers are min–max, box is 25–75th percentile, lines are median values normalized to the geometric mean of Donor 1–3 HPV E6E7. l – n smRNA-ISH images of organoids derived from hCEcto ( l ), hCEcto E6E7 ( m ), and KC-Epi ( n ) probed with HPV16 L1 and E7; Nuclei are shown in blue. Images are representatives of three replicates. Statistical significance was calculated using ( i , j ) two-sided t test, ( k ) One-way ANOVA, P -values are indicated. Source data are provided as a Source Data file.

Article Snippet: Primary cells were expanded in collagen-coated (Sigma, #C3867) tissue culture flasks until they reached 70–80% confluence in human ectocervical primary cell medium (Advanced DMEM/F-12 (Invitrogen, 12634) supplemented with 12 mM HEPES, 1% GlutaMax, 1% B27, 1% N2, 10 ng/mL human EGF (Invitrogen, 15630-056, 35050-038, 17504-044, 17502048, PHG0311), 0.5 μg/mL hydrocortisone (Sigma, H0888-1G), 100 ng/mL human noggin (Peprotech, 120-10 C), 100 ng/mL human FGF-10 (Peprotech, 100-26-25), 1.25 mM N-acetyl-L-cysteine (Sigma, A9165-5G), 2 μM TGF-β receptor kinase Inhibitor IV (SB431542, Calbiochem), 10 μM ROCK inhibitor (Y-27632) (Sigma, Y0503), 10 mM nicotinamide (Sigma, N0636), 10 μM forskolin (Sigma, F6886) before seeding ~20,000 cells/50 μL in Matrigel (BD, #356231) for culturing organoids or for maintenance of 2D stem cells on lethally irradiated J2-3T3 fibroblast feeder cells.

Techniques: Stem Cell Culture, Immunolabeling, Transduction, Luciferase, Control, Expressing, Western Blot, Quantitative RT-PCR, Derivative Assay

a Heatmap of DEG in hCEcto and hCEcto E6E7 2D stem cells with or without Ctr infection; Columns represent biological replicates. b Venn diagram shows genes regulated in hCEcto 2D stem cells after Ctr infection or HPV E6E7 expression. Red box highlights genes upregulated by HPV E6E7 and downregulated by Ctr. c Heatmap showing GSEA enrichment (–log10( P -value)) of genes that share Cis-regulatory motifs for transcription factors. d – g hCEcto and hCEcto E6E7 2D stem cells were infected with Ctr for 48h. mRNA expression of RB1 (d), E2F1 ( e ), and TP53 ( f ) analyzed by qRT-PCR. Data represent mean ± SD from three biological replicates. Statistical significance was calculated using two-sided t test, P -values are indicated. ( g ) Immunoblot analysis for indicated proteins; β-actin. Densitometry values for E2F1 normalized to β-actin; representing FC compared to hCEcto. Data represent three biological replicates. h Dot plot showing up-or down-regulated KEGG pathways among DEG from hCEcto and hCEcto E6E7 2D stem cells with or without Ctr infection. Red arrows highlight DNA repair pathways. Dot diameter refers to the gene ratio within a group. Fill color depicts the adjusted P -value. i Heatmap of genes upregulated in ectocervical stem and differentiated cells from ref. correlated with expression profiles of hCEcto and hCEcto E6E7 organoid with or without Ctr infection. j Heatmap showing expression of genes associated with GO terms for cornification, epidermal cell differentiation, keratinization, skin and epidermis development in hCEcto and hCEcto E6E7 organoids with or without Ctr infection. k Heatmap of DEG from hCEcto E6E7 organoids with or without Ctr infection and their corresponding expression in normal and CIN1-3 tissues. The color bar depicts expression values after subtracting the mean of uninfected hCEcto samples ( i , j ) and normal cervical tissue samples ( k ) and dividing by the SD of each probe. l , m Confocal images of hCEcto 2D stem cells ( l ) and organoids ( m ) infected for 36 h and 5 d, respectively, with Ctr and immunolabelled for Ki67, γH2AX, and Chlamydia MOMP. l , m Data represent three biological replicates. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Modelling Chlamydia and HPV co-infection in patient-derived ectocervix organoids reveals distinct cellular reprogramming

doi: 10.1038/s41467-022-28569-1

Figure Lengend Snippet: a Heatmap of DEG in hCEcto and hCEcto E6E7 2D stem cells with or without Ctr infection; Columns represent biological replicates. b Venn diagram shows genes regulated in hCEcto 2D stem cells after Ctr infection or HPV E6E7 expression. Red box highlights genes upregulated by HPV E6E7 and downregulated by Ctr. c Heatmap showing GSEA enrichment (–log10( P -value)) of genes that share Cis-regulatory motifs for transcription factors. d – g hCEcto and hCEcto E6E7 2D stem cells were infected with Ctr for 48h. mRNA expression of RB1 (d), E2F1 ( e ), and TP53 ( f ) analyzed by qRT-PCR. Data represent mean ± SD from three biological replicates. Statistical significance was calculated using two-sided t test, P -values are indicated. ( g ) Immunoblot analysis for indicated proteins; β-actin. Densitometry values for E2F1 normalized to β-actin; representing FC compared to hCEcto. Data represent three biological replicates. h Dot plot showing up-or down-regulated KEGG pathways among DEG from hCEcto and hCEcto E6E7 2D stem cells with or without Ctr infection. Red arrows highlight DNA repair pathways. Dot diameter refers to the gene ratio within a group. Fill color depicts the adjusted P -value. i Heatmap of genes upregulated in ectocervical stem and differentiated cells from ref. correlated with expression profiles of hCEcto and hCEcto E6E7 organoid with or without Ctr infection. j Heatmap showing expression of genes associated with GO terms for cornification, epidermal cell differentiation, keratinization, skin and epidermis development in hCEcto and hCEcto E6E7 organoids with or without Ctr infection. k Heatmap of DEG from hCEcto E6E7 organoids with or without Ctr infection and their corresponding expression in normal and CIN1-3 tissues. The color bar depicts expression values after subtracting the mean of uninfected hCEcto samples ( i , j ) and normal cervical tissue samples ( k ) and dividing by the SD of each probe. l , m Confocal images of hCEcto 2D stem cells ( l ) and organoids ( m ) infected for 36 h and 5 d, respectively, with Ctr and immunolabelled for Ki67, γH2AX, and Chlamydia MOMP. l , m Data represent three biological replicates. Source data are provided as a Source Data file.

Article Snippet: Primary cells were expanded in collagen-coated (Sigma, #C3867) tissue culture flasks until they reached 70–80% confluence in human ectocervical primary cell medium (Advanced DMEM/F-12 (Invitrogen, 12634) supplemented with 12 mM HEPES, 1% GlutaMax, 1% B27, 1% N2, 10 ng/mL human EGF (Invitrogen, 15630-056, 35050-038, 17504-044, 17502048, PHG0311), 0.5 μg/mL hydrocortisone (Sigma, H0888-1G), 100 ng/mL human noggin (Peprotech, 120-10 C), 100 ng/mL human FGF-10 (Peprotech, 100-26-25), 1.25 mM N-acetyl-L-cysteine (Sigma, A9165-5G), 2 μM TGF-β receptor kinase Inhibitor IV (SB431542, Calbiochem), 10 μM ROCK inhibitor (Y-27632) (Sigma, Y0503), 10 mM nicotinamide (Sigma, N0636), 10 μM forskolin (Sigma, F6886) before seeding ~20,000 cells/50 μL in Matrigel (BD, #356231) for culturing organoids or for maintenance of 2D stem cells on lethally irradiated J2-3T3 fibroblast feeder cells.

Techniques: Infection, Expressing, Quantitative RT-PCR, Western Blot, Cell Differentiation

Cytotoxicity and antiviral activity of Avirulins in female reproductive tract epithelium. ( a – c ) Immortalized epithelial cell lines derived from the vaginal wall, ectocervix, and endocervix (VK2, Ect1, and End1, respectively) were treated with Avirulin lead compounds Av-5, Av-14, and Av-26 at a range of concentrations from 0.78–100 µM for 24 h. Cytotoxicity was measured using a commercially available luminescence-based cytotoxicity assay, CytotoxGlo. Cytotoxicity of treatments was compared to DMSO vehicle control, with the detergent Triton-X 100 for complete cell death. ( n = 4, error = SEM). ( d ) Av-5 was incubated with clarified vaginal fluid at 37 °C for 2 h, then diluted with culture media and applied to TZM-bl. Cells were infected with BaL for 24 h, then inhibition was determined using the luciferase reporter assay. Cervico–vaginal fluid from healthy women is inherently antiviral, as shown in the vehicle control. n = 3, error = SEM * = p > 0.05. ( e ) Cytotoxicity of vaginal fluids and 5 µM Av-5 on TZM-bl measured using CytotoxGlo assay. Cervico–vaginal fluid (CVF) dilutions with Av-5 or equivalent DMSO were incubated at 37 °C for 2 h, then applied to TZM-bl. Cell death at 24 h was determined by subtraction of total cell luminescence from dead cell luminescence. n = 3, error = SEM.

Journal: Viruses

Article Title: Avirulins, a Novel Class of HIV-1 Reverse Transcriptase Inhibitors Effective in the Female Reproductive Tract Mucosa

doi: 10.3390/v11050408

Figure Lengend Snippet: Cytotoxicity and antiviral activity of Avirulins in female reproductive tract epithelium. ( a – c ) Immortalized epithelial cell lines derived from the vaginal wall, ectocervix, and endocervix (VK2, Ect1, and End1, respectively) were treated with Avirulin lead compounds Av-5, Av-14, and Av-26 at a range of concentrations from 0.78–100 µM for 24 h. Cytotoxicity was measured using a commercially available luminescence-based cytotoxicity assay, CytotoxGlo. Cytotoxicity of treatments was compared to DMSO vehicle control, with the detergent Triton-X 100 for complete cell death. ( n = 4, error = SEM). ( d ) Av-5 was incubated with clarified vaginal fluid at 37 °C for 2 h, then diluted with culture media and applied to TZM-bl. Cells were infected with BaL for 24 h, then inhibition was determined using the luciferase reporter assay. Cervico–vaginal fluid from healthy women is inherently antiviral, as shown in the vehicle control. n = 3, error = SEM * = p > 0.05. ( e ) Cytotoxicity of vaginal fluids and 5 µM Av-5 on TZM-bl measured using CytotoxGlo assay. Cervico–vaginal fluid (CVF) dilutions with Av-5 or equivalent DMSO were incubated at 37 °C for 2 h, then applied to TZM-bl. Cell death at 24 h was determined by subtraction of total cell luminescence from dead cell luminescence. n = 3, error = SEM.

Article Snippet: The vaginal, ectocervical, and endocervical epithelium derived cell lines VK2, Ect1, and End1, respectively, were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained with keratinocyte serum free media (KSFM) supplemented with bovine pituitary extract, epidermal growth factor, and calcium chloride in tissue culture treated plates, as per ATCC instructions.

Techniques: Activity Assay, Derivative Assay, Cytotoxicity Assay, Control, Incubation, Infection, Inhibition, Luciferase, Reporter Assay