Journal: Translational Cancer Research

Article Title: Effects of gold nanoparticles on normal hepatocytes in radiation therapy

doi: 10.21037/tcr-21-1855

Figure Lengend Snippet: Effects of AuNPs + radiation combined treatment on the cell proliferation of normal hepatocytes and hepatic stellate cell line. (A) MTT assays were performed using HH cells treated with AuNPs or X-ray irradiation. (B) Transmission electron microscopy images of AuNPs uptake inside the cell. (C) MTT assays were performed using hepatocyte cells (HH: left panel) and hepatic stellate cell line (LX-2: right panel) treated with AuNPs or X-ray irradiation, or a combination of the two treatments. The blue arrows indicate the magnification of AuNPs. MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide; HH, human hepatocyte cell.

Article Snippet: Human hepatocyte cells (HH) were obtained from Sciencell Research Laboratories (Catalog #5200; Carlsbad, CA, USA) and cultured in hepatocyte medium supplemented with 5% fetal bovine serum (FBS), 1% penicillin and streptomycin, and 1% hepatocyte growth supplement (all from Sciencell).

Techniques: Irradiation, Transmission Assay, Electron Microscopy

Journal: Translational Cancer Research

Article Title: Effects of gold nanoparticles on normal hepatocytes in radiation therapy

doi: 10.21037/tcr-21-1855

Figure Lengend Snippet: Cell SF of AuNPs + radiation combined treatment of normal hepatocytes. Clonogenic assay of hepatocytes after treatment with AuNPs or X-ray irradiation or a combination of the two treatments. SF, survival fraction; IR, irradiation.

Article Snippet: Human hepatocyte cells (HH) were obtained from Sciencell Research Laboratories (Catalog #5200; Carlsbad, CA, USA) and cultured in hepatocyte medium supplemented with 5% fetal bovine serum (FBS), 1% penicillin and streptomycin, and 1% hepatocyte growth supplement (all from Sciencell).

Techniques: Clonogenic Assay, Irradiation

Journal: Translational Cancer Research

Article Title: Effects of gold nanoparticles on normal hepatocytes in radiation therapy

doi: 10.21037/tcr-21-1855

Figure Lengend Snippet: Effects on apoptosis of AuNPs + radiation combined treatment of hepatocytes. (A) Cells were treated with PI at 48 h post-treatment after each indicated treatment and analyzed by FACS. (B) The indicated antibodies were used for western blotting of hepatocytes treated with AuNPs or X-ray irradiation or a combination of the two treatments. Values represent the means of 3 experiments ± SD. (C) Cells were treated with AuNPs and/or X-ray irradiation or a combination of the two treatments for 24 h. The cell-cycle distribution of subG1 was analyzed quantitatively. *, P<0.05 and **, P<0.01. IR, irradiation; PARP, poly (ADP-ribose) polymerase; PI, propidium iodide; FACS, fluorescence-activated cell sorting.

Article Snippet: Human hepatocyte cells (HH) were obtained from Sciencell Research Laboratories (Catalog #5200; Carlsbad, CA, USA) and cultured in hepatocyte medium supplemented with 5% fetal bovine serum (FBS), 1% penicillin and streptomycin, and 1% hepatocyte growth supplement (all from Sciencell).

Techniques: Western Blot, Irradiation, Fluorescence, FACS

Journal: Translational Cancer Research

Article Title: Effects of gold nanoparticles on normal hepatocytes in radiation therapy

doi: 10.21037/tcr-21-1855

Figure Lengend Snippet: IL-8 concentrations secreted from normal hepatocytes after AuNPs + X-ray irradiation combined treatment. In all therapeutic groups, a decrease in the amount of IL-8 was observed in the radiation alone group. The combination therapy did not lead to significantly lower levels of IL-8 than radiation alone. IL-8, interleukin-8; IR, irradiation.

Article Snippet: Human hepatocyte cells (HH) were obtained from Sciencell Research Laboratories (Catalog #5200; Carlsbad, CA, USA) and cultured in hepatocyte medium supplemented with 5% fetal bovine serum (FBS), 1% penicillin and streptomycin, and 1% hepatocyte growth supplement (all from Sciencell).

Techniques: Irradiation

Journal: Stem cell research

Article Title: Hepatocyte-like cells differentiated from human induced pluripotent stem cells: Relevance to cellular therapies

doi: 10.1016/j.scr.2012.06.004

Figure Lengend Snippet: Generation of HLCs from hiPSCs. A. Cell morphology by light phase microscopy. The cells gradually transformed into less dense, flatter layers with prominent nuclei (S1), a spiky shape, and then formed an epithelial monolayer (S4). HLCs with large cytoplasmic-to-nuclear ratio, numerous, and prominent nucleoli were observed on day 23, which were of similar morphology and character to primary hepatocytes. B. Relative expression of early stage genes of differentiation. The early stage of hiPSCs differentiation was characterized by the loss of pluripotency marker OCT4. All of the other genes were expressed significantly higher at day 23 (HLCs) than at day 0 (hiPSCs). The special markers for different stages of differentiation (GATA4 for S1, HNF4α for S2, AFP for S3 and Alb for S4) confirmed the successful differentiation of hiPSCs to HLCs. C. Immunofluorescence staining of human cells during 4 stages (23 days) of differentiation. Human hepatocytes are included for comparison (far right column).

Article Snippet: The human hepatocytes were purchased from Yecuris Corporation.

Techniques: Microscopy, Transformation Assay, Expressing, Marker, Immunofluorescence, Staining, Comparison

Journal: Stem cell research

Article Title: Hepatocyte-like cells differentiated from human induced pluripotent stem cells: Relevance to cellular therapies

doi: 10.1016/j.scr.2012.06.004

Figure Lengend Snippet: Characteristics of the HLCs. A. Relative expression of three hepatocyte-related genes using QRT-PCR analyses. Day 23 HLCs expressed higher levels of AFP RNA and lower levels of Alb mRNA than the primary hepatocytes. B. Accumulation of albumin and AFP in the culture supernatant were consistent with expression of these genes in iPS cells (day 0), HLCs (day 23) and hepatocytes. No human albumin was detected in medium of day 0 cultures, which confirmed specificity of our human albumin ELISA. Data are represented as mean ± standard deviation. C. Immunofluorescence staining for AFP and Alb. On day 23 most HLCs stained positively for AFP and Alb, while human hepatocytes only stained positively for Alb.

Article Snippet: The human hepatocytes were purchased from Yecuris Corporation.

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Standard Deviation, Immunofluorescence, Staining

Journal: Stem cell research

Article Title: Hepatocyte-like cells differentiated from human induced pluripotent stem cells: Relevance to cellular therapies

doi: 10.1016/j.scr.2012.06.004

Figure Lengend Snippet: Expression of urea cycle genes and ureagenesis. A. The QRT-PCR assay of urea cycle genes. HLCs expressed lower mRNA level of urea cycle genes than the primary hepatocytes. B. Hepatoblasts (day 18) and HLCs (day 23) showed the ability to produce urea. However, the concentration of urea in these cultures was significantly lower than cultures of primary hepatocytes (p<0.01). Furthermore, the formation of heavy urea from heavy ammonia was only demonstrated by primary hepatocytes, indicating that only primary hepatocytes were capable of normal ureagenesis. Data are represented as mean ± standard deviation.

Article Snippet: The human hepatocytes were purchased from Yecuris Corporation.

Techniques: Expressing, Quantitative RT-PCR, Concentration Assay, Standard Deviation

Journal: Stem cell research

Article Title: Hepatocyte-like cells differentiated from human induced pluripotent stem cells: Relevance to cellular therapies

doi: 10.1016/j.scr.2012.06.004

Figure Lengend Snippet: CYP3A4 activity and diazepam metabolism. A. Expression of CYP3A4 and CYP2C19 increased significantly during the 4 stages of differentiation and was greatest in primary human hepatocytes. ND—expression of CYP3A4 was not detectable in hiPSCs. B. Clearance of diazepam and formation of its three major metabolites (nordiazepam, temazepam, oxazepam) was highest in cultures of primary hepatocytes (p<0.01), indicating greater phase 1 metabolism by primary hepatocyte than HLCs (day 23) or iPSCs (day 0). Data are represented as mean ± standard deviation.

Article Snippet: The human hepatocytes were purchased from Yecuris Corporation.

Techniques: Activity Assay, Expressing, Standard Deviation

Journal: Stem cell research

Article Title: Hepatocyte-like cells differentiated from human induced pluripotent stem cells: Relevance to cellular therapies

doi: 10.1016/j.scr.2012.06.004

Figure Lengend Snippet: Mitochondrial features during differentiation of hiPSCs to hepatocytes. A. Expression of Polg, Polg2, Ucp2, Atp5g1 and Atp8 role steadily during stages of differentiation, while expression of Tfam declined over this time frame. B. The ratio of mitochondrial DNA to nuclear DNA rose significantly (p<0.01) during differentiation reflecting the robust expansion of mitochondria in differentiating cells. ND1 and ND5 are genes encoded on mitochondrial DNA. Data are represented as mean ± standard deviation. C. Top row—cells were examined by confocal microscopy using MitoTracker Red to assess their expanding density of functional mitochondria (red stain) during stages of differentiation. Nuclei were stained blue by DAPI. Middle row (Low power TEM) and Lower Row (high power TEM)—mitochondria were observed to elongate, develop swollen cristae and dense matrices, and migrate deeper into the cytoplasm during stages of differentiation. The mitochondria of HLCs (day 23) appeared less mature than the mitochondria in primary hepatocytes. D. Mitochondria were counted from TEM images. The number of mitochondria rose steadily through stages of differentiation and reached a maximum in primary hepatocytes. The number of mitochondria in HLCs (day 23) was significantly less than in primary hepatocytes (p<0.01). Data are represented as mean ± standard deviation. E. The level of oxygen consumption by HLCs (day 23) was significantly lower than primary hepatocytes (p<0.01). Data are represented as mean ± standard deviation.

Article Snippet: The human hepatocytes were purchased from Yecuris Corporation.

Techniques: Expressing, Standard Deviation, Confocal Microscopy, Functional Assay, Staining

Journal: Stem cell research

Article Title: Hepatocyte-like cells differentiated from human induced pluripotent stem cells: Relevance to cellular therapies

doi: 10.1016/j.scr.2012.06.004

Figure Lengend Snippet: PAS staining for glycogen synthesis. Left—hiPSCs stained very weakly positive for PAS on day 0. Middle—HLCs stained positively for PAS on day 23. A focus of less differentiated cells that do not stain for PAS is shown in the upper center of the image. Primary hepatocytes uniformly stain positive for PAS.

Article Snippet: The human hepatocytes were purchased from Yecuris Corporation.

Techniques: Staining

Journal: PLoS Pathogens

Article Title: Lipoprotein Receptors Redundantly Participate in Entry of Hepatitis C Virus

doi: 10.1371/journal.ppat.1005610

Figure Lengend Snippet: (A) Relative mRNA expression of VLDLR in various tissues was determined using the NextBio Body Atlas application. (B) mRNA expression of SR-B1, LDLR, VLDLR and GAPDH in Huh7 cells and primary human hepatocyte (PHH) were determined by qRT-PCR. Relative expression levels of mRNA were calculated based on the expression level of GAPDH. (C) VLDLR-HA was exogenously expressed in parental, SR-KO, LD-KO and SR/LD-DKO Huh7 cells by infection with lentiviral vectors. Expressions of VLDLR in these cells were determined by immunoblotting analysis (upper panel). Parental, CD81 KO and SR/LD-DKO Huh7 cells expressing SR-B1, LDLR or VLDLR were infected with HCVcc at an MOI of 1 and intracellular HCV RNA levels were determined at 24 h post-infection (lower panel). (D) VLDLR-HA was exogenously expressed in CD81 KO, CLDN1 KO and OCLN KO Huh7 cells by infection with lentiviral vectors. Expressions of VLDLR, CD81, CLDN1 and OCLN in these cells were determined by immunoblotting analysis (upper panel). Cells were infected with HCVcc at an MOI of 1 and intracellular HCV RNA levels were determined at 24 h post-infection by qRT-PCR (lower panel). (E) SR-B1, LDLR and VLDLR were exogenously expressed in SR/LD-DKO Huh7 cells by infection with lentiviral vectors. Cells were infected with Con1-JFH1 or Jc1 at an MOI of 1, and intracellular HCV RNA levels were determined at 24 h post-infection by qRT-PCR. (F) Sera (100μl) from chimeric mice infected with HCV were inoculated into SR/LD-DKO Huh7 cells expressing either SR-B1, LDLR or VLDLR in 24 well plate. Intracellular HCV RNA levels were determined at 72 h post-infection. In all cases, asterisks indicate significant differences (*P<0.05; **P<0.01) versus the results for control cells.

Article Snippet: The primary human hepatocyte (PHH) was purchased from PhoenixBio.

Techniques: Expressing, Quantitative RT-PCR, Infection, Western Blot, Control