hutu80  (ATCC)

Journal: Genes to Cells

Article Title: Establishment of a Novel Caco‐2‐Based Cell Culture System for Human Sapovirus Propagation

doi: 10.1111/gtc.70007

Figure Lengend Snippet: Susceptibility of various cell lines to HuSaV infection. (A) Changes in HuSaV RNA copy numbers in the culture supernatants of HuTu80 and HEK293T cells following inoculation with a HuSaV GI.1 (AH20)‐positive stool suspension (~4 × 10 7 copies/well in 96‐well plates). Each dot represents individual data points; bars indicate the geometric mean value of the HuSaV RNA copy numbers; error bars denote the geometric standard deviation (SD). This experiment was performed once with five technical replicates. (B) Changes in HuSaV RNA copy numbers in the culture supernatants of Caco‐2, HCT15, HCT116, Caco‐2/Cas9, and C2BBe1 cells following inoculation with a HuSaV GI.1 (AH20)‐positive stool suspension (~2 × 10 6 copies/well in 96‐well plates). Each dot represents individual data points; bars indicate the geometric mean HuSaV RNA copy numbers; error bars denote the geometric SD. This experiment was performed once with five technical replicates. (C) Immunofluorescence staining of the viral protein VP1 in HuTu80, HEK293T, Caco‐2, and Caco‐2/Cas9 cells at 3 dpi with a HuSaV GI.1 (AH20)‐positive stool suspension; upper panels: 4× objective lens, lower panels: 40× objective lens. (D) Flow cytometry analysis of Caco‐2 and Caco‐2/Cas9 cells infected with a HuSaV GI.1 (AH20)‐positive stool suspension at 4 dpi. Blue: VP1‐negative cells (uninfected), red: VP1‐positive cells (infected).

Article Snippet: As suggested by Oka et al., successful replication in HuTu80 cells may require highly passaged cells; newly obtained HuTu80 cells from ATCC may not support HuSaV replication (Oka et al. ).

Techniques: Infection, Suspension, Standard Deviation, Immunofluorescence, Staining, Flow Cytometry

Journal: Genes to Cells

Article Title: Establishment of a Novel Caco‐2‐Based Cell Culture System for Human Sapovirus Propagation

doi: 10.1111/gtc.70007

Figure Lengend Snippet: Serial passage of HuSaV GI.1 in HuTu80, HEK293T, Caco‐2, and Caco‐2/Cas9 cells. HuSaV RNA copy numbers in the culture supernatants at 0 or 1 dpi (immediately after medium replacement) and 7 dpi for each passage, up to P + 5, in (A) HuTu80, (B) HEK293T, (C) Caco‐2, and (D) Caco‐2/Cas9 cells.

Article Snippet: As suggested by Oka et al., successful replication in HuTu80 cells may require highly passaged cells; newly obtained HuTu80 cells from ATCC may not support HuSaV replication (Oka et al. ).

Techniques:

Journal: Genes to Cells

Article Title: Establishment of a Novel Caco‐2‐Based Cell Culture System for Human Sapovirus Propagation

doi: 10.1111/gtc.70007

Figure Lengend Snippet: Susceptibility of various cell lines to HuSaV infection. (A) Changes in HuSaV RNA copy numbers in the culture supernatants of HuTu80 and HEK293T cells following inoculation with a HuSaV GI.1 (AH20)‐positive stool suspension (~4 × 10 7 copies/well in 96‐well plates). Each dot represents individual data points; bars indicate the geometric mean value of the HuSaV RNA copy numbers; error bars denote the geometric standard deviation (SD). This experiment was performed once with five technical replicates. (B) Changes in HuSaV RNA copy numbers in the culture supernatants of Caco‐2, HCT15, HCT116, Caco‐2/Cas9, and C2BBe1 cells following inoculation with a HuSaV GI.1 (AH20)‐positive stool suspension (~2 × 10 6 copies/well in 96‐well plates). Each dot represents individual data points; bars indicate the geometric mean HuSaV RNA copy numbers; error bars denote the geometric SD. This experiment was performed once with five technical replicates. (C) Immunofluorescence staining of the viral protein VP1 in HuTu80, HEK293T, Caco‐2, and Caco‐2/Cas9 cells at 3 dpi with a HuSaV GI.1 (AH20)‐positive stool suspension; upper panels: 4× objective lens, lower panels: 40× objective lens. (D) Flow cytometry analysis of Caco‐2 and Caco‐2/Cas9 cells infected with a HuSaV GI.1 (AH20)‐positive stool suspension at 4 dpi. Blue: VP1‐negative cells (uninfected), red: VP1‐positive cells (infected).

Article Snippet: The human duodenal cancer cell line HuTu80 was purchased from ATCC and cultured in Iscove's Modified Dulbecco's Medium (IMDM; SIGMA) supplemented with 1% GlutaMAX‐I (Gibco, Massachusetts, USA), 1% penicillin/streptomycin (Nacalai Tesque), and 5% heat‐inactivated fetal bovine serum (FBS: CAPRICORN, Ebsdorfergrund, Germany).

Techniques: Infection, Suspension, Standard Deviation, Immunofluorescence, Staining, Flow Cytometry

Journal: Genes to Cells

Article Title: Establishment of a Novel Caco‐2‐Based Cell Culture System for Human Sapovirus Propagation

doi: 10.1111/gtc.70007

Figure Lengend Snippet: Serial passage of HuSaV GI.1 in HuTu80, HEK293T, Caco‐2, and Caco‐2/Cas9 cells. HuSaV RNA copy numbers in the culture supernatants at 0 or 1 dpi (immediately after medium replacement) and 7 dpi for each passage, up to P + 5, in (A) HuTu80, (B) HEK293T, (C) Caco‐2, and (D) Caco‐2/Cas9 cells.

Article Snippet: The human duodenal cancer cell line HuTu80 was purchased from ATCC and cultured in Iscove's Modified Dulbecco's Medium (IMDM; SIGMA) supplemented with 1% GlutaMAX‐I (Gibco, Massachusetts, USA), 1% penicillin/streptomycin (Nacalai Tesque), and 5% heat‐inactivated fetal bovine serum (FBS: CAPRICORN, Ebsdorfergrund, Germany).

Techniques: