Batch Cell Culture Techniques
Culture Techniques
Culture Media
Evidence of exponential growth of an anammox population in an anaerobic batch culture. (1/89)
Twenty-five replicates of growth medium for anaerobic ammonium oxidation (anammox) containing (15)N-labeled ammonium and non-labeled nitrite were inoculated into an anammox enrichment culture at low density, and anaerobically incubated batchwise. In the headspace, (29)N(2) partial pressure linearly increased via anammox in 25 vials, confirming that anammox populations were viable in all subcultures. On prolonged incubation, exponential increases in (29)N(2) were not observed in all but 13 subcultures, suggesting that the anammox population may not proliferate unless all conditions for growth are satisfied. The estimated first-order rate coefficients in those 13 subcultures varied from 0.0029 to 0.0048 h(-1). (+info)A novel recirculating flow-perfusion bioreactor for periosteal chondrogenesis. (2/89)
(+info)Biodegradation kinetics of 4-fluorocinnamic acid by a consortium of Arthrobacter and Ralstonia strains. (3/89)
(+info)Physico-chemical characteristics and functional properties of chitin and chitosan produced by Mucor circinelloides using yam bean as substrate. (4/89)
(+info)Evaluation of a laboratory-scale bioreactive in situ sediment cap for the treatment of organic contaminants. (5/89)
(+info)Optimisation of surface expression using the AIDA autotransporter. (6/89)
(+info)Parametric optimization of feruloyl esterase production from Aspergillus terreus strain GA2 isolated from tropical agro-ecosystems cultivating sweet sorghum. (7/89)
A fungal strain, Aspergillus terreus strain GA2, isolated from an agricultural field cultivating sweet sorghum, produced feruloyl esterase using maize bran. In order to obtain maximum yields of feruloyl esterase, the solid state fermentation (SSF) conditions for enzyme production were standardized. Effective feruloyl esterase production was observed with maize bran as substrate followed by wheat bran, coconut husk, and rice husk among the tested agro-waste crop residues. Optimum particle size of 0.71- 0.3 mm and moisture content of 80% favored enzyme production. Moreover, optimum feruloyl esterase production was observed at pH 6.0 and a temperature of 30 degrees C. Supplementation of potato starch (0.6%) as the carbon source and casein (1%) as the nitrogen source favored enzyme production. Furthermore, the culture produced the enzyme after 7 days of incubation when the C:N ratio was 5. Optimization of the SSF conditions revealed that maximum enzyme activity (1,162 U/gds) was observed after 7 days in a production medium of 80% moisture content and pH 6.0 containing 16 g maize bran [25% (w/v)] of particle size of 0.71-0.3 mm, 0.6% potato starch, 3.0% casein, and 64 ml of formulated basal salt solution. Overall, the enzyme production was enhanced by 3.2-fold as compared with un-optimized conditions. (+info)Repeated-batch operation of immobilized beta-galactosidase inclusion bodies-containing Escherichia coli cell reactor for lactose hydrolysis. (8/89)
In this study, we investigated the performance of an immobilized beta-galactosidase inclusion bodies-containing Escherichia coli cell reactor, where the cells were immobilized in alginate beads, which were then used in repeated-batch operations for the hydrolysis of o-nitrophenyl-beta-D-galactoside or lactose over the long-term. In particular, in the Tris buffer system, disintegration of the alginate beads was not observed during the operation, which was observed for the phosphate buffer system. The o-nitrophenyl-beta-D-galactoside hydrolysis was operated successfully up to about 80 h, and the runs were successfully repeated at least eight times. In addition, hydrolysis of lactose was successfully carried out up to 240 h. Using Western blotting analyses, it was verified that the beta-galactosidase inclusion bodies were sustained in the alginate beads during the repeated-batch operations. Consequently, we experimentally verified that beta-galactosidase inclusion bodies-containing Escherichia coli cells could be used in a repeated-batch reactor as a biocatalyst for the hydrolysis of o-nitrophenyl-beta-D-galactoside or lactose. It is probable that this approach can be applied to enzymatic synthesis reactions for other biotechnology applications, particularly reactions that require long-term and stable operation. (+info)Batch cell culture techniques refer to a method of growing cells in which all the necessary nutrients are added to the culture medium at the beginning of the growth period. The cells are allowed to grow and multiply until they exhaust the available nutrients, after which the culture is discarded. This technique is relatively simple and inexpensive but lacks the ability to continuously produce cells over an extended period.
In batch cell culture, cells are grown in a closed system with a fixed volume of medium, and no additional nutrients or fresh medium are added during the growth phase. The cells consume the available nutrients as they grow, leading to a decrease in pH, accumulation of waste products, and depletion of essential factors required for cell growth. As a result, the cells eventually stop growing and enter a stationary phase, after which they begin to die due to lack of nutrients and buildup of toxic metabolites.
Batch cell culture techniques are commonly used in research settings where large quantities of cells are needed for experiments or analysis. However, this method is not suitable for the production of therapeutic proteins or other biologics that require continuous cell growth and protein production over an extended period. For these applications, more complex culture methods such as fed-batch or perfusion culture techniques are used.
Cell culture is a technique used in scientific research to grow and maintain cells from plants, animals, or humans in a controlled environment outside of their original organism. This environment typically consists of a sterile container called a cell culture flask or plate, and a nutrient-rich liquid medium that provides the necessary components for the cells' growth and survival, such as amino acids, vitamins, minerals, and hormones.
There are several different types of cell culture techniques used in research, including:
1. Adherent cell culture: In this technique, cells are grown on a flat surface, such as the bottom of a tissue culture dish or flask. The cells attach to the surface and spread out, forming a monolayer that can be observed and manipulated under a microscope.
2. Suspension cell culture: In suspension culture, cells are grown in liquid medium without any attachment to a solid surface. These cells remain suspended in the medium and can be agitated or mixed to ensure even distribution of nutrients.
3. Organoid culture: Organoids are three-dimensional structures that resemble miniature organs and are grown from stem cells or other progenitor cells. They can be used to study organ development, disease processes, and drug responses.
4. Co-culture: In co-culture, two or more different types of cells are grown together in the same culture dish or flask. This technique is used to study cell-cell interactions and communication.
5. Conditioned medium culture: In this technique, cells are grown in a medium that has been conditioned by previous cultures of other cells. The conditioned medium contains factors secreted by the previous cells that can influence the growth and behavior of the new cells.
Cell culture techniques are widely used in biomedical research to study cellular processes, develop drugs, test toxicity, and investigate disease mechanisms. However, it is important to note that cell cultures may not always accurately represent the behavior of cells in a living organism, and results from cell culture experiments should be validated using other methods.
Culture techniques are methods used in microbiology to grow and multiply microorganisms, such as bacteria, fungi, or viruses, in a controlled laboratory environment. These techniques allow for the isolation, identification, and study of specific microorganisms, which is essential for diagnostic purposes, research, and development of medical treatments.
The most common culture technique involves inoculating a sterile growth medium with a sample suspected to contain microorganisms. The growth medium can be solid or liquid and contains nutrients that support the growth of the microorganisms. Common solid growth media include agar plates, while liquid growth media are used for broth cultures.
Once inoculated, the growth medium is incubated at a temperature that favors the growth of the microorganisms being studied. During incubation, the microorganisms multiply and form visible colonies on the solid growth medium or turbid growth in the liquid growth medium. The size, shape, color, and other characteristics of the colonies can provide important clues about the identity of the microorganism.
Other culture techniques include selective and differential media, which are designed to inhibit the growth of certain types of microorganisms while promoting the growth of others, allowing for the isolation and identification of specific pathogens. Enrichment cultures involve adding specific nutrients or factors to a sample to promote the growth of a particular type of microorganism.
Overall, culture techniques are essential tools in microbiology and play a critical role in medical diagnostics, research, and public health.
Bacteriological techniques refer to the various methods and procedures used in the laboratory for the cultivation, identification, and study of bacteria. These techniques are essential in fields such as medicine, biotechnology, and research. Here are some common bacteriological techniques:
1. **Sterilization**: This is a process that eliminates or kills all forms of life, including bacteria, viruses, fungi, and spores. Common sterilization methods include autoclaving (using steam under pressure), dry heat (in an oven), chemical sterilants, and radiation.
2. **Aseptic Technique**: This refers to practices used to prevent contamination of sterile materials or environments with microorganisms. It includes the use of sterile equipment, gloves, and lab coats, as well as techniques such as flaming, alcohol swabbing, and using aseptic transfer devices.
3. **Media Preparation**: This involves the preparation of nutrient-rich substances that support bacterial growth. There are various types of media, including solid (agar), liquid (broth), and semi-solid (e.g., stab agar). The choice of medium depends on the type of bacteria being cultured and the purpose of the investigation.
4. **Inoculation**: This is the process of introducing a bacterial culture into a medium. It can be done using a loop, swab, or needle. The inoculum should be taken from a pure culture to avoid contamination.
5. **Incubation**: After inoculation, the bacteria are allowed to grow under controlled conditions of temperature, humidity, and atmospheric composition. This process is called incubation.
6. **Staining and Microscopy**: Bacteria are too small to be seen with the naked eye. Therefore, they need to be stained and observed under a microscope. Gram staining is a common method used to differentiate between two major groups of bacteria based on their cell wall composition.
7. **Biochemical Tests**: These are tests used to identify specific bacterial species based on their biochemical characteristics, such as their ability to ferment certain sugars, produce particular enzymes, or resist certain antibiotics.
8. **Molecular Techniques**: Advanced techniques like PCR and DNA sequencing can provide more precise identification of bacteria. They can also be used for genetic analysis and epidemiological studies.
Remember, handling microorganisms requires careful attention to biosafety procedures to prevent accidental infection or environmental contamination.
Culture media is a substance that is used to support the growth of microorganisms or cells in an artificial environment, such as a petri dish or test tube. It typically contains nutrients and other factors that are necessary for the growth and survival of the organisms being cultured. There are many different types of culture media, each with its own specific formulation and intended use. Some common examples include blood agar, which is used to culture bacteria; Sabouraud dextrose agar, which is used to culture fungi; and Eagle's minimum essential medium, which is used to culture animal cells.