Protein response of insect cells to bioreactor environmental stresses

Huhtala, Anne; Linko, P.; Mutharasan, Raj

doi:10.1016/j.jbiotec.2005.05.009

Cited by 9 publications

(16 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the present study, the expression level of HSP90-1 was downregulated and then upregulated, but remained lower compared to the initial level. It is possible that the 4 ppt was so low that it fell beyond the salinity range that induces the production of HSP90-1; thus, disrupting protein synthesis in organisms (Huhtala et al, 2005). A similar expression pattern has been reported in the gill of Crassostrea hongkongensis under low salinity stress (Fu et al, 2011).…”

Section: Discussionsupporting

confidence: 56%

mRNA expression profiles of heat shock proteins of wild and salinity-tolerant swimming crabs, Portunus trituberculatus, subjected to low salinity stress

Bao

Zhang

et al. 2014

Genet. Mol. Res.

View full text Add to dashboard Cite

ABSTRACT. Challenged by the low salinity, 4 parts per thousand (4 ppt), for 72 h, the survivals of swimming crabs (Portunus trituberculatus) were collected as the screened group (SG, tolerant to low salinity). Aiming at identifying the mechanism of low salinity tolerance, quantitative real-time PCR was employed to investigate the expression profiles of 4 HSP genes (HSP60, HSP70, HSP90-1, HSP90-2) in the hepatopancreas of wild (WG) and screened (SG) groups of P. trituberculatus exposed to low salinity (4 ppt). The results showed that 3 of the candidate genes (HSP60, HSP70, HSP90-1) exhibited similarly downregulated expression profiles in the first 3 h (P < 0.05), which became upregulated from 3 h to 72 h after being subjected to low salinity conditions. In contrast, the expression profile of the HSP90-2 gene was upregulated during the first 6 h for the WG, and during the first 12 h for the SG, after which it became downregulated. HSP90-1 and HSP90-2 were highly expressed at 12 h after low salinity challenge in the SG, but not the WG. The response of these 2 genes to salinity stress indicates their suitability as biomarkers to differentiate SG from WG crabs. The results indicate that HSP genes are involved in the adaptation of crabs to low salinity exposure, and that different HSPs have diverse functions in response to low salinity stress in P. trituberculatus. In addition, HSP expression in SG indicates that this group is more tolerant to low salinity conditions compared to WG.

show abstract

Section: Discussionsupporting

confidence: 56%

mRNA expression profiles of heat shock proteins of wild and salinity-tolerant swimming crabs, Portunus trituberculatus, subjected to low salinity stress

Bao

Zhang

et al. 2014

Genet. Mol. Res.

View full text Add to dashboard Cite

show abstract

“…Moreover, the results of the expression obtained in T. castaneum cells are consistent with the results of the work of Huhtala et al . (), where protein response to different environmental stress conditions was studied in Spodoptera frugiperda cells. That paper showed that the cells began to express higher level of heat shock proteins than controls at temperatures over 39 °C.…”

Section: Discussionmentioning

confidence: 94%

“…Huhtala et al . () exposed S. frugiperda cells to small pH variations over long time and high variations over short periods finding in both cases no change in viability and protein production of the cells, which seems to be consistent with the results obtained in this study. The difference in gene expression between Hsp27 and Hsp68a in this work is significant, but is closer to reference levels than temperature and UV samples.…”

Section: Discussionmentioning

confidence: 98%

“…Huhtala et al . () detected in S. frugiperda protein production up to 45 °C but viability decreased progressively after 42 °C.…”

Section: Discussionmentioning

confidence: 99%

“…Huhtala et al . () studied the production of heat shock proteins in Spodoptera frugiperda cells in response to heat shock, salinity, pH changes, alcohol, and oxygen deprivation conditions. Coleoptera cell lines are increasingly being used on different studies; Anthonomus grandis (McIntosh et al ., ), Leptinotarsa decemlineata (Lynn, ; Long et al ., ), Xylotrechus pyrrhoderus (Iwabuchi, ), Plagionotus christophi (Hoshino et al ., ), or Blaps rhynchoptera (Zhang et al ., ).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Real‐time cell analysis and heat shock protein gene expression in the TcA Tribolium castaneum cell line in response to environmental stress conditions

et al. 2016

View full text Add to dashboard Cite

The rust red flour beetle, Tribolium castaneum (Herbst, 1797) (Coleoptera: Tenebrionidae), is a pest of stored grain and one of the most studied insect model species. Some of the previous studies involved heat response studies in terms of survival and heat shock protein expression, which are regulated to protect other proteins against environmental stress conditions. In the present study, we characterize the impedance profile with the xCELLigence Real-Time Cell Analyzer and study the effect of increased temperature in cell growth and viability in the cell line BCIRL-TcA-CLG1 (TcA) of T. castaneum. This novel system measures cells behavior in real time and is applied for the first time to insect cells. Additionally, cells are exposed to heat shock, increased salinity, acidic pH and UV-A light with the aim of measuring the expression levels of Hsp27, Hsp68a, and Hsp83 genes. Results show a high thermotolerance of TcA in terms of cell growth and viability. This result is likely related to gene expression results in which a significant up-regulation of all studied Hsp genes is observed after 1 h of exposure to 40 °C and UV light. All 3 genes show similar expression patterns, but Hsp27 seems to be the most affected. The results of this study validate the RTCA method and reveal the utility of insect cell lines, real-time analysis and gene expression studies to better understand the physiological response of insect cells, with potential applications in different fields of biology such as conservation biology and pest management.

show abstract

Improving AAV vector yield in insect cells by modulating the temperature after infection

Aucoin

Perrier

Kamen

2007

Biotech & Bioengineering

View full text Add to dashboard Cite

Vectors based on adeno-associated viruses (AAV) are sought for therapeutic gene delivery because of their ability to transduce a variety of tissues with no significant immunological response. Production using the baculovirus expression vector (BEV)/insect cell system has the potential to meet the needs for pre-clinical and clinical trials. In this co-infection system, three baculoviruses are used to produce the AAV vector. A strategy aimed at increasing encapsidation/maturation of the viral vector involved varying the temperature over the course of the process. Cultures were subjected to temperature changes at various times pre- and post-infection (up to 24 h post-infection). It was found that raising the culture temperature to 30 degrees C at the time of infection nearly tripled the infectious titer. In fact, increasing the temperature to 30 degrees C at any time in the process investigated resulted in an increase in titer. Also, raising the culture to 33 degrees C or lowering the temperature to 24 degrees or 21 degrees C resulted in lower titers. The rise in infectious titer was also confirmed by an increase in DNase resistant particles (DRPs). Varying the temperature, however, did not affect the total amount of capsids significantly. Therefore increasing the culture temperature resulted in better encapsidation as determined by the ratio of capsids to DRPs to infectious particles. It is believed that an increase in early proteins and possibly a quicker cascade of baculovirus infection events resulted in this increased packaging efficiency.

show abstract

Protein response of insect cells to bioreactor environmental stresses

Cited by 9 publications

References 30 publications

mRNA expression profiles of heat shock proteins of wild and salinity-tolerant swimming crabs, Portunus trituberculatus, subjected to low salinity stress

mRNA expression profiles of heat shock proteins of wild and salinity-tolerant swimming crabs, Portunus trituberculatus, subjected to low salinity stress

Real‐time cell analysis and heat shock protein gene expression in the TcA Tribolium castaneum cell line in response to environmental stress conditions

Improving AAV vector yield in insect cells by modulating the temperature after infection

Contact Info

Product

Resources

About