Serum is commonly used as a supplement to cell culture media. It provides a broad spectrum of macromolecules, carrier proteins for lipoid substances and trace elements, attachment and spreading factors, low molecular weight nutrients, and hormones and growth factors. The most widely used animal serum supplement is fetal bovine serum, FBS. Since serum in general is an ill-defined component in cell culture media, a number of chemically defined serum-free media formulations have been developed in the last two decades. Besides modern cell biological advances in cell and tissue culture and efforts towards a standardisation of cell culture protocols in Good Cell Culture Practice, in addition, considerable ethical concerns were raised recently about the harvest and collection of fetal bovine serum. Thus, in order to decrease the annual need for bovine fetuses in terms of the 3Rs through any reduction in the use or partial replacement of serum, as well as in terms of an improvement of cell and tissue culture methodology, serum-free cell culture represents a modern, valuable and scientifically well accepted alternative to the use of FBS in cell and tissue culture.

Alternatives to the use of fetal bovine serum: serum-free cell culture.

Fetal bovine serum (FBS) is a ubiquitously used essential supplement in cell culture media. However, there are serious scientific and ethical concerns about the use of FBS regarding its harvest and production. During the last three decades, FBS could be substituted by other supplements or by the use of defined chemical components in serum-free cell culture. A number of serum-free medium formulations have been described for mammalian and insect cell lines as well as for primary cultures. However, the switch to serum-free media still demands a time-consuming literature survey and a manufacturer search for appropriate medium formulations, respectively. Here we present the second collection of commercially available serum-free media in an updated, freely accessible interactive online database. Searches for serum-free media and continuous cell lines already adapted to serum-free culture can be performed according to various criteria. These include the degree of chemical definition, e.g. serum-free (SF), animal-derived component free (ADCF) or chemically defined (CD), and the type of medium, e.g. basal media, medium supplements, or full replacement media. In order to specify the cell lines that are adapted to serum-free media, search terms like species, organ, tissue, cell type and disease can be used. All commercially available serum-free media and adapted cell lines currently available from major distributors (e.g. ATCC, ECACC and DMSZ) are included in the database. Despite an extensive search for serum-free media and adapted cell lines, detailed information from certain companies and suppliers is still lacking and is specifically highlighted. It is intended to create a platform for the interactive exchange of information and experience by experts in the field in order to continuously improve and extend the serum-free online database. The database is accessible at http://www.goodcellculture.com/

/pdf/serum-free-cell-culture-the-serum-free-media-interactive-m8igwrno7s.pdf

Serum-free cell culture: the serum-free media interactive online database.

Growth factors and antimicrobial factors of bovine colostrum

Fibronectins

We measured the concentrations of protein, insulin, and epidermal growth factor (EGF) in human milk from mothers delivering at term. Samples were obtained from d 1 (colostrum) to d 42 after birth. Human colostrum contains very high concentrations of protein [83.7 +/- 7.4 g/l (SEM)], insulin (3.75 +/- 0.88 nM), and EGF (53.9 +/- 6.9 nM). Similar concentrations have been measured in prebirth milk. Insulin, EGF, and protein in milk decline rapidly during the first few days of lactation but remain constant thereafter. Although the concentrations of insulin and EGF in mature milk are only 10% of those in colostrum, they are considerably higher than in serum. We also showed that human milk has a growth-promoting activity in cultured cells, causing a stimulation of protein synthesis in L6 myoblasts and 3T3-L1 fibroblasts and an increase in DNA synthesis in L6 cells and T47D breast cancer cells. This mitogenic activity declines as lactation progresses, with a similar time-course to the fall in insulin and EGF; however, the cell lines used here are not responsive to either of these two growth factors in the range of concentrations found in milk. This indicates that human milk also contains high concentrations of additional, unidentified growth factors. The occurrence of high concentrations of growth factors in milk suggests that they may be important for the proliferation and differentiation of infant tissues.

Changes in the Growth-Promoting Activity of Human Milk during Lactation

Seven established cell lines, including both epithelial cells and fibroblasts (MDCK, Vero, CV-1, NRK, 3T3, F2408, and NIL8) and four early passage cell strains (bovine articular chondrocytes, bovine smooth muscle cells, human foreskin fibroblasts, and rat embryo cells) were cultured in serum-free medium supplemented with milk obtained 1 day after birth (colostrum) or 80 days after birth (older milk). MDCK, Vero, CV-1, NRK, and 3T3 grew readily in colostrum and attained saturation densities ranging from 22% to 63% of that in serum. There was no growth of F2408, NIL8, or the early passage strains in bovine colostrum. None of the 11 cell cultures grew in older milk. The temporal dependence of growth in milk was examined in detail using MDCK cells. Growth equivalent to that in serum occurred in 3% colostrum and in 15% milk obtained 2 days after birth. Milk obtained 3 days and 10 days after birth was not effective as a growth supplement for MDCK cells at any concentration. Those cells, unable to grow in colostrum or in older milk, could be induced to grow if culture dishes were precoated with fibronectin. In addition to fibronectin, it was necessary in some cultures to supplement colostrum or older milk with insulin and/or transferrin in order to achieve growth. In the presence of fibronectin and appropriate factors, the final saturation density attained in colostrum or older milk ranged from 25% to 100% of that in serum. The fibronectin contents of bovine colostrum and milk were determined. The fibronectin level of colostrum was found to be approximately 5% of bovine serum. There was no detectable fibronectin in the 80-day-old milk.

The serum‐free growth of cultured cells in bovine colostrum and in milk obtained later in the lactation period

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The serum‐free growth of cultured cells in bovine colostrum and in milk obtained later in the lactation period