Rat carcinoma cells in long-term, serum-free culture provide a continuing supply of collagenase

1985 ◽  
Vol 5 (12) ◽  
pp. 1071-1077 ◽  
Author(s):  
Geoffrey A. Stevenson ◽  
J. Guy Lyons ◽  
David A. Cameron ◽  
Robert L. O'Grady

Neoplastic, epithelial cells derived from a spontaneously-arising rat mammary carcinoma have been cultured in a defined medium, in the absence of serum, continuously, for over 2 years. The medium is a mixture of Ham's F12 and Dulbecco's Modified Eagle's media supplemented with insulin, transferrin and bovine serum albumin. The cells have retained their potential to produce tumours and, in culture, a true vertebrate collagenase. This system provides a continuing supply of vertebrate collagenase through the application of recently developed methods.

Metallomics ◽  
2010 ◽  
Vol 2 (3) ◽  
pp. 204-210 ◽  
Author(s):  
Lourdes Garza-Ocañas ◽  
Domingo A. Ferrer ◽  
Justin Burt ◽  
Luis A. Diaz-Torres ◽  
Mónica Ramírez Cabrera ◽  
...  

1980 ◽  
Vol 94 (4) ◽  
pp. 571-576 ◽  
Author(s):  
Kirsti Käpyaho

Abstract. Rat ovarian ornithine decarboxylase activity could be stimulated in vitro by a variety of factors, which apparently have different modes of action. Ovarian cells prepared from pre-pubertal rats by collagenase dispersion exhibited a low but detectable ornithine decarboxylase activity after a 6-h incubation in a defined medium. The enzyme activity was markedly enhanced in vitro by hCG, which also produced increased accumulation of cyclic AMP and stimulated the secretion of progesterone. In addition to the gonadotrophin, ovarian ornithine decarboxylase activity was strikingly stimulated by some non-essential amino acids, and especially by bovine serum albumin. While markedly enhancing ornithine decarboxylase activity, none of the latter additions increased the accumulation of cyclic AMP or enhanced the secretion of progesterone. Bovine serum albumin enhanced powerfully ornithine decarboxylase activity in vitro at very small concentrations (from 0.75 μm). The half-life of the enzyme remained unchanged (26—28 min) upon stimulation indicating that the stimulation mechanism did not involve any stabilization of the enzyme.


1980 ◽  
Vol 87 (2) ◽  
pp. 434-441 ◽  
Author(s):  
R A Wolfe ◽  
G H Sato ◽  
D B McClure

In this communication we describe serum-free culture conditions for the serial propagation of the C6 glioma cell line. The growth rate, saturation density, and morphology of these cells are equivalent to those of their serum-grown counterparts when cultured in a 3:1 mixture of Dulbecco's modified Eagle's medium and Ham's medium F-12 supplemented with trace elements, insulin, transferrin, fibroblast growth factor, linoleic acid complexed to fatty acid-free bovine serum albumin, and a serum-spreading factor (SSF) partially purified from human plasma. The requirement for SSF in the medium can be satisfied by preincubating the tissue culture dishes with SSF. Tissue culture dishes sequentially pretreated with poly-D-lysine and purified cold insouluble globulin will also substitute for this requirement. The fatty acid-free bovine serum albumin/linoleic acid complex increases the growth rate of these cells but has no appreciable effect on their morphology, saturation density, or ability to grow with repeated subculture. The growth stimulation caused by this complex appears to be dependent on the fatty acid, as the fatty acid-free bovine serum albumin alone has no effect on the growth rate. Linoleic acid is cytotoxic in the absence of bovine serum albumin, and the fatty acid-free bovine serum albumin prevents this toxicity. Other fatty acids including oleic, arachidonic, and palmitic only partially substitute for the growth-promoting effect of linoleic acid.


2005 ◽  
Vol 17 (8) ◽  
pp. 815 ◽  
Author(s):  
A. Reis ◽  
G. J. McCallum ◽  
T. G. McEvoy

Sheep zygotes were cultured in serum-free or serum-supplemented media to determine effects on blastocyst yields and within-blastocyst abundance and distribution of neutral lipid droplets. Embryos cultured in synthetic oviduct fluid supplemented with bovine serum albumin (0.4% w/v) (SBSA) generated similar blastocyst yields (mean ± s.e.m. = 20% ± 5) to those in synthetic oviduct fluid supplemented with serum (10% v/v) from ewes fed a diet containing 0% (SZFO; 26% ± 2) or 3% fish oil (S3FO; 23% ± 3). SBSA zygotes generated more good-quality blastocysts than their SZFO or S3FO counterparts (P < 0.05). Within-blastocyst abundance of neutral lipid droplets was non-uniform; data were collected from discrete embryo sectors (each = 2700 µm2) representing highest (H), intermediate (I) and lowest (L) densities of accumulation. For all sectors, area (µm2) occupied by lipid droplets in SBSA blastocysts (mean H = 470; I = 370; L = 245) was smaller (P < 0.01) than occupied in others (SBSA : SZFO = 1 : 1.41, 1 : 1.48 and 1 : 1.42; SBSA : S3FO = 1 : 1.36, 1 : 1.30 and 1 : 1.31; data for H, I and L, respectively). Among S3FO blastocysts only, inferior quality was associated with greater lipid abundance. Overall, embryo culture in the presence of serum increased neutral lipid droplet abundance but accumulation was non-uniform.


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