Journal: Engineering in Life Sciences

Article Title: Overexpression of GRP78 enhances survival of CHO cells in response to serum deprivation and oxidative stress

doi: 10.1002/elsc.201500152

Figure Lengend Snippet: Viability of Chinese hamster ovary (CHO) cells in response to serum deprivation and oxidative stress. CHO cells were seeded in 12‐well plates at a density of 1.5 × 105 cells per well and grown for 24 h. Then cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium, serum free medium with or without 200 μM H2O2. In order to ensure that other nutrient limitations were not experienced, cells were fed fresh medium every 48 h. Cells were taken every 2 days and analyzed for cell viability using Annexin V/PI double staining. Error bars represent SD for each measurement.

Article Snippet: The Chinese hamster ovary (CHO) cells, human stomach adenocarcinoma (AGS) cell line, and human hepatoma cell line HepG2 (China Center for Type Culture Collection, Wuhan, P. R. China) were cultured in Roswell Park Memorial Institute (RPMI) 1640 and DMEM medium (Gbico BRL, NY, USA) supplemented with 10% FBS (Sijiqing, Hangzhou, China) and 100 U/mL ampicillin, 100 mg/mL streptomycin in an incubator containing 5% CO 2 at 37°C (standard conditions).

Techniques: Cell Culture, Double Staining

Journal: Engineering in Life Sciences

Article Title: Overexpression of GRP78 enhances survival of CHO cells in response to serum deprivation and oxidative stress

doi: 10.1002/elsc.201500152

Figure Lengend Snippet: Glucose‐regulated protein 78 (GRP78) engineering protected Chinese hamster ovary (CHO) cells against injury induced by H2O2. (A) Cells synchronized in serum‐free medium were treated with H2O2 at increasing concentration for 4 h and then cultured with 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT; 5 mg/mL) for 4 h. Bar graph represented the percentage of viable cells assayed by MTT. (B) The level of cleaved caspase‐3 in H2O2‐treated cells was detected by western blot (upper). The lower left bar graph represented densitometric analysis of GRP78 levels of the western blots and the lower right of cleaved caspase‐3. Data were mean values ± SD, n > 3.*p < 0.05,**p < 0.01,***p < 0.001, NS, nonsignificant difference versus negative control.

Article Snippet: The Chinese hamster ovary (CHO) cells, human stomach adenocarcinoma (AGS) cell line, and human hepatoma cell line HepG2 (China Center for Type Culture Collection, Wuhan, P. R. China) were cultured in Roswell Park Memorial Institute (RPMI) 1640 and DMEM medium (Gbico BRL, NY, USA) supplemented with 10% FBS (Sijiqing, Hangzhou, China) and 100 U/mL ampicillin, 100 mg/mL streptomycin in an incubator containing 5% CO 2 at 37°C (standard conditions).

Techniques: Concentration Assay, Cell Culture, Western Blot, Negative Control

Journal: Engineering in Life Sciences

Article Title: Overexpression of GRP78 enhances survival of CHO cells in response to serum deprivation and oxidative stress

doi: 10.1002/elsc.201500152

Figure Lengend Snippet: Glucose‐regulated protein 78 (GRP78) engineering enhanced yields of TfR‐Ab and improved cell viability. Chinese hamster ovary (CHO) and CHO modified by GRP78 (CHO‐GRP78) cells were seeded in 12‐well plates at a density of 2 × 105 cells per well and transiently transfected with pOptiVEC™‐TOPO®/TfR‐Ab, followed by culturing with SFM4CHO™ medium. (A) The concentration of TfR‐Ab in the supernatant was detected by ELISA assay. (B) The quality of TfR‐Ab in the supernatant was evaluated by its binding ability with TfR+ HepG2 cells using flow cytometry (FCM). The bar graphs represented FCM analysis of mean fluorescence index (MFI). (C) Bar graph represented FCM analysis of the percentage viability of CHO and CHO‐GRP78 cells. (D) The viable cell numbers of CHO and CHO‐GRP78 cells were counted after Trypan blue staining. Data were mean values ± SD, n > 3.*p < 0.05,**p < 0.01,***p < 0.001 versus negative control.

Article Snippet: The Chinese hamster ovary (CHO) cells, human stomach adenocarcinoma (AGS) cell line, and human hepatoma cell line HepG2 (China Center for Type Culture Collection, Wuhan, P. R. China) were cultured in Roswell Park Memorial Institute (RPMI) 1640 and DMEM medium (Gbico BRL, NY, USA) supplemented with 10% FBS (Sijiqing, Hangzhou, China) and 100 U/mL ampicillin, 100 mg/mL streptomycin in an incubator containing 5% CO 2 at 37°C (standard conditions).

Techniques: Modification, Transfection, Concentration Assay, Enzyme-linked Immunosorbent Assay, Binding Assay, Flow Cytometry, Fluorescence, Staining, Negative Control