Search PubMedSearch

Biomedical subjects

B Griffiths

Publications and source records attributed to B Griffiths.

11 recordsLinked to original sources

Alternative strategies to the scaleup of animal cells.

The use of porous microcarriers has given manufacturers the option of having a high cell density/medium volume (10-100 L)/high specific productivity/long-term process. The advantages of the system are listed in Table 7 and many of the criteria listed in Table 1 are met. The question of being able to license products from continuous processes has been addressed and guidelines are being prepared. Methods are available to compare the master and end-of-process (extended) cell banks to ensure that no change has occurred to the cell or product. Rigid attention to process definition and validation, as well as to the definition of production lots, overcomes many of the other reservations. In conclusion, the method has been demonstrated as technically feasible, economically advantageous, and licensable and the many advantages of a high density perfusion process (listed in Table 8) make porous microcarrier technology an ideal scaleup process because it is a simple, reliable, and scalable system.

Animal Population Groups

DNA fingerprinting--a valuable new technique for the characterisation of cell lines.

DNA fingerprinting is an important new development for the authentication of cell lines. Multilocus methods such as those developed by Alec Jeffreys provide information on a wide range of genetic loci throughout the human genome and thus give a useful genetic "snap-shot" of a cell culture. Our work has shown that Jeffreys multilocus fingerprinting method can be applied to cell lines from a wide range of animals including reptiles, birds, fish and diverse mammals. It can also differentiate very closely related cell lines including those from the same mouse strain. Routine fingerprint analysis has enabled an unprecedented level of confidence in the consistency of cell stocks. Our results demonstrate that this straightforward method represents a powerful and readily interpreted system for cell authentication and exclusion of cross-contamination.

Animals

Differential mucin gene expression in human pancreatic and colon cancer cells.

The purpose of this study was to determine the quantity and nature of the mucins synthesized and secreted by four different pancreatic cancer cell lines. Well- to moderately-differentiated SW1990 and CAPAN-2 human pancreatic cancer cells were found to produce more high-Mr glycoprotein (HMG) than less-differentiated MIA PaCa-2 and PANC-1 cells. Most of the labelled HMG was secreted within 24 h. The results of chemical and enzymic degradation, ion-exchange chromatography and density-gradient centrifugation indicated that the HMG in SW1990 and CAPAN-2 cells has the properties expected for mucins, whereas much of the HMG in MIA PaCa-2 and PANC-1 cells may not be mucin, but proteoglycan. These results are consistent with immunoblots and Northern blots showing the presence of apomucin and apomucin mRNA in SW1990 and CAPAN-2 cells, but not in MIA PaCa-2 and PANC-1 cells. The Western blots and Northern blots also show that SW1990 and CAPAN-2 cells, like breast cancer cells, have the mammary-type apomucin and mRNA coded by the MUC1 gene, but lack the intestinal type apomucin and mRNA coded by the MUC2 gene. In contrast, the colon cancer cell lines tested in culture express apomucin and mRNA coded by MUC2 but not by MUC1.

Adenocarcinoma

ELFT: a gene that directs the expression of an ELAM-1 ligand.

The LECCAMs are a family of cell adhesion molecules implicated in certain inflammatory processes. ELAM-1, a LECCAM found on the surface of activated endothelial cells, can mediate adhesion of neutrophils, monocytes, and certain cell lines to endothelial cells in vitro. No ligand for any LECCAM has yet been fully characterized. Here we report the cloning of a cDNA, ELFT (ELAM-1 ligand fucosyltransferase), that can confer ELAM-1 binding activity when transfected into nonbinding cell lines. ELFT encodes a 46 kd protein that has alpha(1,3)fucosyltransferase activity, suggesting that a fucosylated carbohydrate structure is an essential component of the ELAM-1 ligand. Furthermore, ELFT is expressed specifically in cell types that bind to ELAM-1, suggesting that this enzyme is an important regulator of inflammatory events in vivo.

Amino Acid Sequence

Assignment of the polymorphic intestinal mucin gene (MUC2) to chromosome 11p15.

A cDNA coding for a mucin expressed in intestine has recently been cloned (Gum et al. 1989). We describe here the use of this cDNA to map the gene (MUC2) to human chromosome 11 using somatic cell hybrids, and to make the regional localization to 11p15 by in situ hybridization. Analysis of the CEPH (Centre d'Etude du Polymorphisme Humain) families revealed that MUC2 forms part of the tight linkage group on 11p15 which contains HRAS, INS, TH and HBBC.

Animals