Rapid Identification of Chinese Hamster Ovary Cell Apoptosis and Its Potential Role in Process Robustness Assessment.

Currently, the assessment of process robustness is often time-consuming, labor-intensive, and material-intensive using process characterization studies. Therefore, a simple and time-saving method is highly needed for the biopharmaceutical industry. Apoptosis is responsible for 80% of Chinese hamster ovary (CHO) cell deaths and affects the robustness of the cell culture process. This study's results showed that a more robust process can support cells to tolerate apoptosis for a longer time, suggesting that the robustness of the process could be judged by the ability of cells to resist apoptosis. Therefore, it is necessary to establish a rapid method to detect the apoptosis of CHO cells. In trying to establish a new method for detecting apoptosis in large-scale cell cultures, glucose withdrawal was studied, and the results showed that CHO cells began to apoptose after glucose was consumed. Then, the concentration of extracellular potassium increased, and a prolongation of apoptosis time was observed. Further study results showed that the process with poor robustness was associated with a higher proportion of apoptosis and extracellular potassium concentration, so potassium could be used as a biochemical index of apoptosis. The strategy we present may be used to expedite the assessment of process robustness to obtain a robust cell culture process for other biologics.

Toxicity assessment of Chlorella vulgaris and Chlorella protothecoides following exposure to Pb(II).

The short- and long-term toxic effects of Pb(II) exposure on Chlorella vulgaris (C. vulgaris) and Chlorella protothecoides (C. protothecoides) were not well understood. The lab study was performed to observe the Pb(II) exposure induced changes. Results of the observations show: (1) higher level of Pb(II) (50 or 80mgL(-1)) could significantly inhibit the growth and chlorophyll a synthesis of both algae in almost all the treatments and dose-response relationships could be clearly observed, (2) the range of EC50 values (24-120h, 67.73-172.45mgL(-1)) indicated that Pb(II) had a relatively limited short-term toxicity to the two algae, while long-term tests (7-28d, 50.41-63.91mgL(-1)) displayed higher toxicity and (3) SOD and CAT activities of both algae after exposed to medium level of Pb(II) were significantly promoted, and their response might be more susceptible in short-term exposure. This research provides a basic understanding of Pb(II) toxicity to aquatic organisms.

Toxicity assessment of zebrafish following exposure to CdTe QDs.

CdTe quantum dots (QDs) are nanocrystals of unique composition and properties that have found many new commercial applications; therefore, their potential toxicity to aquatic organisms has become a hot research topic. The lab study was performed to determine the developmental and behavioral toxicities to zebrafish under continuous exposure to low concentrations of CdTe QDs (1-400 nM) coated with thioglycolic acid (TGA). The results show: (1) the 120 h LC(50) of 185.9 nM, (2) the lower hatch rate and body length, more malformations, and less heart beat and swimming speed of the exposed zebrafish, (3) the brief burst and a higher basal swimming rate of the exposed zebrafish larvae during a rapid transition from light-to-dark, and (4) the vascular hyperplasia, vascular bifurcation, vascular crossing and turbulence of the exposed FLI-1 transgenic zebrafish larvae.