Embryo cells cell preparation

Several attenuated strains have been developed for use in vaccine preparations. The most commonly used is the Jeryl Linn strain, which is propagated in chick embryo cell culture. This vaccine has been administered to well over 50 million people worldwide and, typically, results in seroconversion rates of over 97 per cent. The Sabin (oral poliomyelitis) vaccine consists of an aqueous suspension of poliomyelitis virus, usually grown in cultures of monkey kidney tissue. It contains approximately 1 million particles of poliomyelitis strains 1,2 or 3 or a combination of all three strains. 

As already indicated, control points in the regulation of the rate of glyeosylation of proteins may be (a) the availability of Dol-P (Section 11,1,a), (b) the D-glucosylation of the lipid-linked oligosaccharide, or its transfer to protein, or both (Section 1,2,a), and, as already discussed, (c) the metabolic fate of GDP-Man. Furthermore, GDP-Man inhibits fonnation of Glc-P-Dol in cell-free preparations from liver,157 and activates formation of GlcNAe-PP-Dol in cell-free preparations from tissues of chick embryo.156... 

The EF cells are prepared from 14-16-d-old embryos (from 14-16 d postcoitus pregnant mice). Since most of the EF cells used for ES cells are under... 

Bunyak, E.B. Hilleman, M.R. Weiber, R.E. Stokes, J., Jr. Live attenuated rubella virus vaccines prepared in duck embryo cell culture. I. Development and clinical testing. J. Am. Med. Assoc. 1968, 204, 195-200. 

Normal animal tissues (e.g., skin, kidney, liver) or whole embryos are commonly used to establish primary cell cultures. To prepare tissue cells for a primary culture, the cell-cell and cell-matrix interactions must be broken. To do so, tissue fragments are treated with a combination of a protease (e.g., trypsin or the collagen-hydrolyzing enzyme collagenase or both) and a divalent cation chelator (e.g., EDTA) that depletes the medium of usable Ca or Mg. The released cells are then placed in dishes in a nutrient-rich, serum-supplemented medium, where they can adhere to the surface and one another. The same protease/chelator solution is used to remove adherent cells from a culture dish for biochemical studies or subculturing (transfer to another dish). 

Ulitzur, N.. and Gruenbaum, Y. (1989). Nuclear envelope assembly around sperm chromatin in cell-free preparations from Drosophila embryos. FEBS Lett. 259,113-116. 

Chicken embryo fibroblast cells (CEF) were prepared from the carcasses of 10-day-old embryos and maintained in complete growth medium as described [5]. The cells were used between the third and seventh subcultivation after initiation of the cultures. Cells were labeled in vivo with 125 juCi of [2,8,5 - H]adenosine per ml of culture media (50.5 Ci mmol 1 Ci = 3.7 x 10 Bq New England Nuclear) as described [5]. 

Syrian hamster embryo and mouse CSH lOTl/2 cells were used for transformation assays. Hamster cells were prepared and grown in Dulbecco s modified Eagle s medium supplemented with 10% fetal calf serum, penicillin (50 U ml" ), and streptomycin (50 jug ml" GIBCO), as previously described [4]. 

The enzyme which isomerizes cycloeucalenol (8-F) to obtusifoliol (9-F), that is, opens the cyclopropane ring, has been demonstrated in cell-free preparations from tissue cultures of bramble (Heintz and Benveniste, 1974) and microsomes of Zea mays embryos the maize enzyme is specific for cycloeucalenol (9-F) (Rahier et al., 1976, 1977). The fact that neither cycloar-... 

Cultures of muscle cells were prepared from superficial pecto-rails muscles of 12- to 13-day-old chicken embryos as indicated in a previous report (Martonosi et al., 1976). The cultures were harvested between 2 and 13 days. The collected muscle cells were repeatedly... 

Historically, primary cultures of hamster embryo cells that served as a source of target cells in the clonal test system were prepared as needed for each experiment. However, variations in susceptibility of the cells prepared from different batches of pooled embryo cells hindered efforts to standardize the system for routine use as a prescreen for carcinogens. This led some investigators to the use of cryopreserved aliquot samples of pooled embryo cells that were... 

In this series of bioassay experiments, several carcinogens failed to transform certain batches of hamster embryo cells, presumably because of a lack of endogenous enzymes required for the metabolic activation of these compounds. In short-term bacterial and mammalian cell mutagenesis systems, these enzymes are routinely provided by the addition of rat liver homogenates. As mentioned earlier, diethylnitrosamine and urethane, which failed to transform hamster embryo cells directly, were activated in a host-mediated in vivo-in vitro cell transformation assay in which the chemicals were inoculated intraperitoneally. In our laboratory, when A -2-acetylaminofluorene, 4-aminoazobenzene, auramine, diethylnitrosamine, 3-methoxy-4-aminoazoben-zene, Natulan, 2-nitrofluorene, p-rosaniline, and urethane were tested in the presence of hamster liver homogenates and appropriate cofactors, all except 4-aminoazobenzene gave positive results. The liver homogenates were prepared from hamsters that were not treated with enzyme inducers such as Aroclor 1254 or phenobarbital. 

Actively growing or secreting tissues, such as microorganisms, young oocytes, differentiating embryos, root tips, liver, nerve, and exocrine cells, all have a high RNA content on the other hand, tissues which produce protein at a slower rate, such as heart, muscle, and kidney, for example, have a lower RNA content. Within a ven cell the rate of protein synthesis is greatest in those subcellular structures richest in RNA (cf. Section II) and in some broken cell preparations (87, 374) there seemed to be a direct correlation between RNA content, or the amount of RNA added, and ability to incorporate amino acids into protein. 

The incorporation of C Meucine into a virus-specific protein, the so-called S-antigen of fowl plague virus, in homogenates of chick embryo cells has been investigated by Mueller et al. 37). The activity of the homogenate (which must be prepared from suitably infected fibroblast cells) is partially sensitive to ribonuclease and is somewhat depressed by the omission of ATP and an ATP-generating system. The addition of rather lai e quantities of embryonic nucleic acid is also required, but the activity of the latter is not destroyed by either ribo- or deoxyribonuclease, or by treatment with alkali. 

Figure 14.3. High-magnification views of organelles from cells prepared by HPF-FS. (a) Microfilaments (arrow) in cross section. Bar, 50 nm. (b) Microfilaments in longitudinal section. The arrows point out areas of apparent transverse elements in the bundle. Bar, 50 nm. (c) Membranes between adjacent cells in a postgastrulation embryo. Bar, 200 nm. (d) Membranes between cells in a pregastrulation embryo. Bar, 200 nm. (e-g) Centrioles from syncytial blastoderm mitotic spindle poles. Bars (e) 200 nm (f,g) 100 nm. See text for additional details.

Embryo Collection and Preparation for Cell Culture, 304 Recording, 304... 

Definition of a toxicity with culture of the primerely trepsinised chicken fibroblasts (TCF) included the following stages - incubation of chicken embryo within 11 day - tripsinisation and cultivation of a TCF cells monolayer - 2 days - contact of the tested preparation with the TCF cells monolayer an and the count of a toxicity by the citopathogenic action (CPA)-1-5 day. Hence, definition of a toxicity on cell culture borrowed 14-18 days and at the last two stages demanded strict sterility. 

Enzymic transfer of D-xylose from uridine 5 -(D-xylopyranosyI-HC pyrophosphate) to L-serine residues of endogenous protein acceptors from (a) a cell tumor of the mouse188 and (b) chick-embryo cartilage189 occurs in cell-free extracts of both of these tissues, in the absence of biosynthesis of protein. The enzyme preparations employed were from the supernatant liquor, although activity was also present in the insoluble fractions. In these two types of tissue, the acceptors are heparin and chondroitin sulfate, respectively, but the presence of other D-xylose-containing glycoproteins in ascites fluid from... 

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