An actively mixed mini-bioreactor for protein production from suspended animal cells

2008 ◽  
Vol 100 (1) ◽  
pp. 72-81 ◽  
Author(s):  
Jinpian Diao ◽  
Lincoln Young ◽  
Peng Zhou ◽  
Michael Louis Shuler
Keyword(s):  
Protein Production ◽  
Animal Cells

Use of regulated secretion in protein production from animal cells: An evaluation with the AtT-20 model cell line

1990 ◽  
Vol 35 (8) ◽  
pp. 771-780 ◽  
Author(s):  
A. Sambanis ◽  
Gregory Stephanopoulos ◽  
A. J. Sinskey ◽  
H. F. Lodish
Keyword(s):  
Cell Line ◽  
Protein Production ◽  
Animal Cells ◽  
Regulated Secretion ◽  
Model Cell

Development of PCL-PEG nanofibrous mats as alternative carriers for recombinant Chinese hamster ovary cells

2012 ◽  
Vol 32 (6-7) ◽  
pp. 453-461
Author(s):  
Mehdi Shakibaie ◽  
Marieh Ghafari Rahbar ◽  
Fatemeh Tabandeh ◽  
Hossein Tavanai ◽  
Alireza Zomorodipour ◽  
...  
Keyword(s):  
Chinese Hamster Ovary ◽  
Chemical Structure ◽  
Protein Production ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Animal Cells ◽  
Ovary Cells ◽  
Mass Cultivation ◽  
Cell Densities ◽  
Nanofiber Mats

Abstract In this study, six polyblended nanofiber mats, composed of poly ε-caprolactone (PCL)-polyethylene glycol (PEG) mixed at different proportions in an electrospinning solution were fabricated, and then utilized alongside commercial polyester microcarriers for the mass cultivation of recombinant Chinese hamster ovary (rCHO) cells. The SEM micrographs showed that the rCHO cells were attached to the nanofiber mats with significant differences in their affinities, which can be related to the chemical structure and hydrophilicity of the nanofiber mats. The cell densities of the rCHO cells grown on the polyblended PCL-PEG nanofiber mat (80:20)14% were much higher when compared with the commercial polyester microcarrier. Moreover, recombinant protein production increased by approximately 20% when cells were grown on the PCL-PEG nanofiber mat (80:20)14%. The results of this research indicated a potential for the replacement of the commercial polyester microcarrier with the PCL-PEG polyblended nanofiber mats, in order to achieve mass cultivation of recombinant animal cells in industrial bioreactors.

Surface Structure of Nucleated Animal Cells: Carbon Replicas

1967 ◽  
Vol 25 ◽  
pp. 120-121
Author(s):  
Robert M. Glaeser ◽  
Thea B. Scott
Keyword(s):  
Cell Surface ◽  
Surface Structure ◽  
Particle Diameter ◽  
Cellular Functions ◽  
Animal Cells ◽  
Replica Technique ◽  
Carbon Replica ◽  
Characteristic Particle ◽  
Cell Surface Structure ◽  
Carbon Replicas

The carbon-replica technique can be used to obtain information about cell-surface structure that cannot ordinarily be obtained by thin-section techniques. Mammalian erythrocytes have been studied by the replica technique and they appear to be characterized by a pebbly or “plaqued“ surface texture. The characteristic “particle” diameter is about 200 Å to 400 Å. We have now extended our observations on cell-surface structure to chicken and frog erythrocytes, which possess a broad range of cellular functions, and to normal rat lymphocytes and mouse ascites tumor cells, which are capable of cell division. In these experiments fresh cells were washed in Eagle's Minimum Essential Medium Salt Solution (for suspension cultures) and one volume of a 10% cell suspension was added to one volume of 2% OsO4 or 5% gluteraldehyde in 0.067 M phosphate buffer, pH 7.3. Carbon replicas were obtained by a technique similar to that employed by Glaeser et al. Figure 1 shows an electron micrograph of a carbon replica made from a chicken erythrocyte, and Figure 2 shows an enlarged portion of the same cell.

Nuclear Envelope Dynamics During Mitosis

1988 ◽  
Vol 46 ◽  
pp. 224-225
Author(s):  
Brian Burke
Keyword(s):  
Nuclear Envelope ◽  
Membrane Vesicles ◽  
Nuclear Pore ◽  
Nuclear Pore Complexes ◽  
Animal Cells ◽  
Interphase Chromosome ◽  
Lamin B ◽  
Chromosome Architecture ◽  
Complex Membrane ◽  
Pore Complexes

The nuclear envelope is a complex membrane structure that forms the boundary of the nuclear compartment in eukaryotes. It regulates the passage of macromolecules between the two compartments and may be important for organizing interphase chromosome architecture. In interphase animal cells it forms a remarkably stable structure consisting of a double membrane ouerlying a protein meshwork or lamina and penetrated by nuclear pore complexes. The latter form the channels for nucleocytoplasmic exchange of macromolecules, At the onset of mitosis, however, it rapidly disassembles, the membranes fragment to yield small vesicles and the lamina, which is composed of predominantly three polypeptides, lamins R, B and C (MW approx. 74, 68 and 65 kDa respectiuely), breaks down. Lamins B and C are dispersed as monomers throughout the mitotic cytoplasm, while lamin B remains associated with the nuclear membrane vesicles.

Localization of Adenosine Triphosphatase Activity in the Phleom of Nicotiana Tabacum

1972 ◽  
Vol 30 ◽  
pp. 230-231
Author(s):  
James Cronshaw ◽  
Jamison E. Gilder
Keyword(s):  
Plasma Membrane ◽  
Atpase Activity ◽  
Adenosine Triphosphatase ◽  
Higher Plants ◽  
High Surface ◽  
Root Tip ◽  
Animal Cells ◽  
Physiological Processes ◽  
Tip Cells ◽  
Atpase Localization

Adenosine triphosphatase (ATPase) activity has been shown to be associated with numerous physiological processes in both plants and animal cells. Biochemical studies have shown that in higher plants ATPase activity is high in cell wall preparations and is associated with the plasma membrane, nuclei, mitochondria, chloroplasts and lysosomes. However, there have been only a few ATPase localization studies of higher plants at the electron microscope level. Poux (1967) demonstrated ATPase activity associated with most cellular organelles in the protoderm cells of Cucumis roots. Hall (1971) has demonstrated ATPase activity in root tip cells of Zea mays. There was high surface activity largely associated with the plasma membrane and plasmodesmata. ATPase activity was also demonstrated in mitochondria, dictyosomes, endoplasmic reticulum and plastids.

Structural aspects of tracheary element differentiation in suspension cultures of Zinnia elegans

1989 ◽  
Vol 47 ◽  
pp. 768-769
Author(s):  
C. H. Haigler ◽  
A. W. Roberts
Keyword(s):  
Tracheary Element ◽  
Vesicle Fusion ◽  
Zinnia Elegans ◽  
Cellulose Synthesis ◽  
Tracheary Elements ◽  
Animal Cells ◽  
Mesophyll Cells ◽  
Fixation Techniques ◽  
Localized Patterns ◽  
Structural Aspects

Tracheary elements, the water-conducting cells in plants, are characterized by their reinforced walls that became thickened in localized patterns during differentiation (Fig. 1). The synthesis of this localized wall involves abundant secretion of Golgi vesicles that export preformed matrix polysaccharides and putative proteins involved in cellulose synthesis. Since the cells are not growing, some kind of endocytotic process must also occur. Many researchers have commented on where exocytosis occurs in relation to the thickenings (for example, see), but they based their interpretations on chemical fixation techniques that are not likely to provide reliable information about rapid processes such as vesicle fusion. We have used rapid freezing to more accurately assess patterns of vesicle fusion in tracheary elements. We have also determined the localization of calcium, which is known to regulate vesicle fusion in plant and animal cells.Mesophyll cells were obtained from immature first leaves of Zinnia elegans var. Envy (Park Seed Co., Greenwood, S.C.) and cultured as described previously with the following exceptions: (a) concentration of benzylaminopurine in the culture medium was reduced to 0.2 mg/l and myoinositol was eliminated; and (b) 1.75ml cultures were incubated in 22 x 90mm shell vials with 112rpm rotary shaking. Cells that were actively involved in differentiation were harvested and frozen in solidifying Freon as described previously. Fractures occurred preferentially at the cell/planchet interface, which allowed us to find some excellently-preserved cells in the replicas. Other differentiating cells were incubated for 20-30 min in 10(μM CTC (Sigma), an antibiotic that fluoresces in the presence of membrane-sequestered calcium. They were observed in an Olympus BH-2 microscope equipped for epi-fluorescence (violet filter package and additional Zeiss KP560 barrier filter to block chlorophyll autofluorescence).

Three-Dimensional reconstruction of isolated centrosomes by automated electron tomography

1994 ◽  
Vol 52 ◽  
pp. 310-311
Author(s):  
M.B. Braunfeld ◽  
M. Moritz ◽  
B.M. Alberts ◽  
J.W. Sedat ◽  
D.A. Agard
Keyword(s):  
Electron Tomography ◽  
Three Dimensional ◽  
Vital Role ◽  
Complex Nature ◽  
Animal Cells ◽  
Microtubule Organizing Center ◽  
Nucleation Sites ◽  
Dimensional Reconstruction ◽  
Organizing Center ◽  
Resolution Model

In animal cells, the centrosome functions as the primary microtubule organizing center (MTOC). As such the centrosome plays a vital role in determining a cell's shape, migration, and perhaps most importantly, its division. Despite the obvious importance of this organelle little is known about centrosomal regulation, duplication, or how it nucleates microtubules. Furthermore, no high resolution model for centrosomal structure exists.We have used automated electron tomography, and reconstruction techniques in an attempt to better understand the complex nature of the centrosome. Additionally we hope to identify nucleation sites for microtubule growth.Centrosomes were isolated from early Drosophila embryos. Briefly, after large organelles and debris from homogenized embryos were pelleted, the resulting supernatant was separated on a sucrose velocity gradient. Fractions were collected and assayed for centrosome-mediated microtubule -nucleating activity by incubating with fluorescently-labeled tubulin subunits. The resulting microtubule asters were then spun onto coverslips and viewed by fluorescence microscopy.

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