Bioreactors : animal cell culture control for bioprocess engineering
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Bioreactor is a key device during in vitro animal cell culture. The stirred-tank bioreactor is one of the most commonly used types, and is used both for industrial applications and laboratory research. Important improvements have been made in the design of traditional bioreactors, and new types of bioreactor are also being developed such as Couette-Taylor bioreactor, multifunctional-membrane bioreactor, and shaking bioreactor. Work is also progressing on techniques to improve the performance of bioreactors, including perfusion culture, the use of microcarriers, and methods of suppressing apoptosis and of monitoring cell growth in real time.
Given the demand for the production by animal cells for use in the growing number of clinical applications, further advances in bioreactor technology can be expected during the next few years. Bioreactors: Animal Cell Culture Control for Bioprocess Engineering presents the design, fabrication, and control of a new type of bioreactor meant especially for animal cell line culture. It reviews the bioreactors particularly designed for the animal cell culture, and addresses the mechanical loads on the cultured cells.
Animal cell processes are primarily used for the production of proteins and viral vaccines of relatively high value. Recent advancements in animal cell bioreactor are reviewed. Summary of time taken for adaptation to serum free in CHO cells using P21 and bcl2 technology.
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The use of continuous chemostat culture which is widely used in microbial cultures provides an opportunity to comprehensively and efficiently survey the limiting factors of growth and productivity under different conditions and to estimate the relative levels of specific key proteins involved in the control of growth and death in sub-optimal nutrient levels hopefully leading to the development of improved and stable process. This approach should readily lead to improvements in the present state of the art for production of recombinant products.
Ex vivo expansion of erythroid progenitors from cord blood and other sources is a potential source for supply of blood products which would significantly improve the transfusion capacity of blood centres Neildez-Nguyen et al. In vitro generated RBCs have all the characteristics of functional, native, adult RBCs in terms of enzyme content, membrane deformability and the capacity to fix and release oxygen.
In this project, we are seeking to develop a continuously perfused free suspension or encapsulated system to provide suitable environment for high growth rate with high potential for fully maturation into RBCs and facilitates long term in vitro growth of haematopoietic stem cells and their differentiation into a pure erythroid lineage. Why is this problem significant? During the last decade there has been an explosion of new knowledge and techniques in the field of haematopoiesis.
Ex vivo expansion of bone marrow cells has been proposed as an effective method for the early recovery from pancytopenia in patients. Most recently, development of novel models of human erythropoiesis that result in mature RBCs promises to herald a new era for the transfusion of red blood cells and to address the pathophysiological abnormalities in congenital RBC diseases and to test the potential of reversing these problems by gene therapy Malik et al. We have found in animal cells used for the production of biopharmaceuticals grown in various systems batch, fed batch and perfusion that:.
We have extensively used flow cytometry to obtain detailed knowledge of the metabolic and synthetic state of the cell population and to understand cell culture bioreaction processes as well as to provide terms for adequate and sensitive process design and control. We currently use the genomic, proteomic and cytomic examination as a critical step toward understanding cellular function and product productivity when cells are grown in serum free media at high cell density culture and when any of the above genes are over-expressed.
The ultimate objective is to advance our molecular and engineering understanding of in vitro culture to solve important problems in cell culture to optimise cell productivity genomics, proteomics and metabolomics.
With use of single cell analytical systems the necessary knowledge can be obtained from multilevel single cell molecular analysis in combination with extensive bioinformatic knowledge extraction cytomics. The advancement in high throughput techniques offers new opportunities for animal cell culture industry. Thanks to SFI, we have now started a unique research programme which will unravel the key characteristics of the cell lines: NS0 myeloma and CHO and develop the necessary new methods and tools.
Bioreactors for animal cell suspension culture
Damaged articular cartilage has a limited ability to repair itself due to the absence of vascularization and nerve endings in the tissue. Traditional therapies to repair damaged cartilage include alloplastic and allogenic implants and more recently autologous chondrocyte transplantation. The former therapies are limited by donor site morbidity, while the latter therapy requires surgical removal of healthy cartilage and is limited by the size of the defect. As an alternative to these current therapies, efforts in tissue engineering of cartilage have led to the development of biocompatible, biodegradable scaffolds onto which chondrocytes are seeded.
One of the challenges that tissue engineers will have to address in the near future is the development of feasible large-scale cell expansion processes. The estimated number of articular cartilage incidences worldwide is around 30 million cases of knee osteoarthritis and 1. If tissue engineering products are to be used for the treatment of these incidences, it is crucial to estimate the scale of cell production needed for all these repair procedures, but such essential question that is rarely approached in reported tissue engineering studies. The table below gives an idea of the dimensional aspect of articular cartilage repair and the estimated cell necessities associated.
Routine tissue culturing methodologies can hardly cope with the scale of cell production required for these procedures. Expansion of cell population in vitro has become an essential step in the process of tissue engineering of articular cartilage, and optimization of the culture conditions and expansion protocols are fundamental issues that need to be addressed. We have established the culture conditions and operation modes to optimise cell expansion in a serial 2D monolayer and 3D culture passaging process.
For this aim, we have found the use of mathematical expressions to define the growth curve of the cultures to be a valuable tool. Currently our aim is to tissue engineer cartilage that can be used for biological repair of damaged articular cartilage or degenerative joint disease. In order to do this cartilage will be engineered that is more similar to natural tissue than current in vitro engineered cartilage. An important issue to address is that of nutrient diffusion limited thickness of engineered cartilage. We propose to develop a novel bioreactor to culture tissue engineered cartilage constructs under hydrodynamic loading and to test the hypothesis that mechanical stimulation to tissue specimens mediates the effectiveness of 3-D scaffolds for cartilage tissue engineering.
Cartilage in human joints is exposed continually to mechanical loading, which is considered to be essential for its homeostasis, and for achieving functional tissue engineered cartilage repair in 3-D biodegradable scaffolds. The cultivation of chondrocytes in polymer scaffolds leads to implants that may potentially be used to repair damaged joint cartilage or for reconstructive surgery Saini and Wick, , Biotechnol Prog. For this technique to be medically applicable the physical parameters that govern cell growth and differentiation in the polymer scaffold must be understood and optimised.
Bioreactors : animal cell culture control for bioprocess engineering
The former therapies are limited by donor site morbidity, while the latter therapy requires surgical removal of healthy cartilage and is limited by the size of the defect Cancedda et al. Sports Med. Tissue engineering of cartilaginous constructs has become an essential methodology to provide functional grafts for the repair of cartilage defects, and both optimisation of the culture conditions and the validation of a production process are fundamental problems that need to be addressed Chaudhuri and Al-Rubeai, Bioreactors for Tissue Engineering, , Springer.
The Cluster aims to develop the scientific and technological knowledge needed for present and future manufacturing applications using plasmas, with a specific emphasis on nano-scale products, process reliability, manufacturing costs and advanced materials processing. It enables the fundamental understanding of plasma processing to be coupled with real word product application in the medical device and food packaging industries. As part of this project, our group will investigate:. Scanning electron microscopy image showing osteoblast adhesion onto plasma modified hydroxyapatite. Carroll S.
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