Primary culture

Product extraction Effluent and waste disposal Medium preparation Seed vessel Purification Cell free supernatant Cell biomass Production bioreactor Downstream processing Medium sterilisation Primary culture Upstream processing... 

Primary cultured cerebellar granule cells Increase in cytosolic [Ca2+] SR 48692 ... 

Animal cell cultures that are initiated from cells removed directly from the animal are called primary cultures (Figure 2). Primary cultures include both explant cultures (i.e., cultures initiated from small pieces of intact tissue), as well as cultures initiated from preparations of individual or dispersed cells (obtained from intact tissue by mechanical or proteolytic dismption). Nerve fiber explant cultures in blood plasma were among the earliest types of tissue cultures (Harrison, 1907). Cells grow out from such tissue explants and form a single layer of cells completely filling the tissue culture vessel surface. Such cell cultures are called confluent monolayers. Confluent monolayers can then be treated with trypsin, so as to remove the individual cells from the culture vessel surface. The resulting cell suspension is then transferred into other culture containers, so that more viable monolayer... 

Figure 2. Initiation of primary cultures and their establishment as cell lines. Primary cultures are initiated from tissue removed from animals. Following multiple subcul-turings, cell lines in some cases are established.

A good example of the problems encountered with the use of serum with primary cultures is illustrated by the case with cultured kidney cells. The kidney epithelial cell line MDCK grows in serum-free medium supplemented with five supplements ... 

Figure 10. Primary cultures of mouse kidney cells. Primary cultures of kidney epithelial cells derived from 10-day-old mice were grown either in hormonally defined medium with five supplements (5 pg/ml insulin, 5 pg/ml transferrin, 25 ng/ml PCE, 5 X10" M hydrocortisone, and 5 x 10" M Tj), or in medium supplemented with 10% fetal calf serum. After 10 days, primary cultures still were epithelial in morphology serum free (a) but were overgrown with fibroblasts with serum (b). (Taub et al., 1979 with permission.)...

Taub, M. Sato, G. (1980). Growth of functional primary cultures of kidney epithelial cells in defined medium. J. Cell. Physiol. 105, 369-378. 

Pomes A, Rodriguez-Farre E, Sunol C. 1994. Disruption of GABA-dependent chloride flux by cyclodienes and hexachlorocyclohexanes in primary cultures of cortical neurons. J Pharmacol Exp Ther 271(3) 1616-1623. 

Rosa R, Rodriguez-Farre E, Sanfeliu C. 1996. Cytotoxicity of hexachlorocyclohexane isomers and cyclodienes in primary cultures of cerebellar granule cells. J Pharmacol Exp Ther 278(1) 163-169. 

Pesonen, M., Goksoyr, A., and Andersson, T. (1992). Expression of P450 lAl in a primary culture of rainbow trout microsomes exposed to B-naphthoflavone or 2,3,7,8-TCDD. Archives of Biochemistry and Biophysics 292, 228-233. 

Vaillant, C., Monod, G., and Volataire, Y. et al. (1989). Measurement and induction of cytochrome P450 and monooxygenases in a primary culture of rainbow trout hepatocytes. Comptes Redus de L Academic des Sciences 308, 83-88. 

The first report of the action of a chemokine on neurons was published in 1993. The study demonstrated that IL-8 could increase the survival of cultured neurons (Araujo and Cotman, 1993). However, as can be appreciated from its name, IL-8 was not known to be a chemokine at that time and was instead classed as an interleukin. Indeed, the expression of chemokine receptors by neurons was not generally appreciated until around 1997/1998 when several reports suggested this. These reports included observations of the expression of chemokine receptors by neuronal cell lines (Hesselgesser et al. 1997), primary cultures of neurons (Meucci et al. 1998 Ohtani et al. 1998), and in brain sections from HlV-1, Alzheimer s disease, and other patients (Horuk et al. 1997 Westmoreland et al. 1998 Xia et al. 1997). Furthermore, data were obtained, suggesting functions for chemokine signaling in the development of the nervous system (Zou et al. 1998) as well as in neuronal survival and communication (Giovannelli et al. 1998 Meucci et al. 1998). 

Secondary cell cultures, which can be prepared by taking cells from some types of primary culture, usually those derived from embryonic tissue, dispersing them by treatment with trypsin and inoculating some into a fresh batch of medium. A limited number of subcultures can be performed with these sorts of cells, up to a maximum of about 50 before the cells degenerate. 

Shanker G, Mutkus LA, Walker SJ, Aschner M. 2002. Methylmercury enhances arachidonic acid release and cytosolic phospholipase A2 expression in primary cultures of neonatal astrocytes. Brain Res Mol Brain Res 106 1-11. 

The phenothiazines, chlorpromazine and promethazine, have been described as inhibitors of CCU-induced lipid peroxidation at relatively high concentrations in rat liver microsomes (Slater, 1968). Structural modifications of chlorpromazine were undertaken to try to increase antioxidant activity and maintain molecular lipophilicity. The 2-N-N-dimethyl ethanamine methanesulphonate-substituted phenothiazine (3) was found to be a potent inhibitor of iron-dependent lipid peroxidation. It was also found to block Cu -catalysed oxidation of LDL more effectively than probucol and to protect primary cultures of rat hippocampal neurons against hydrogen peroxide-induced toxicity in vitro (Yu et al., 1992). 

Chapin RE, Phelps JL, Burka LT, et al. 1991. The effects of tri-ozt/zo-cresyl phosphate and metabolites on rat Sertoli cell function in primary culture. Toxicol Appl Pharmacol 108 194-208. 

Cullen CF, Milner MJ 1991 Parameters of growth in primary cultures and cell lines established from Drosophila imaginal discs. Tissue Cell 23 29—39 Davis KT, Shearn A 1977 In vitro growth of imaginal disks from Drosophila melanogaster. Science... 

Figure 14 Observed permeability coefficients of urea and mannitol across monolayers of rat alveolar epithelial cells in primary culture in the Transwell system are correlated with transepithelial electrical resistance and days in culture.

WS Marshall, JW Hanrahan. (1991). Anion channels in the apical membrane of mammalian corneal epithelium primary cultures. Invest Ophthalmol Vis Sci 32 1562-1568. 

SK Basu, IS Haworth, MB Bolger, VHL Lee. (1998). Proton-driven dipeptide uptake in primary cultured rabbit conjunctival epithehal cells. Invest Ophthalmol Vis Sci 39 2365-2373. 

Pienta RJ, Poiley JA, Lebherz WB III. 1977. Morphological transformation of early-passage golden Syrian hamster embryo cells derived from cryopreserved primary cultures as a reliable in vitro bioassay for identifying diverse carcinogens. Int J Cancer 19 642-655. 

Peehl, DM. 2004. Are primary cultures realistic models for prostate cancer J Cell Biochem 91 185-195. 

Gilad GM, Gilad VH (1986) Cytotoxic effects of monodansylcadaverina and methylamine in primary cultures of rat cerebellar neurons. Int J Dev Neurosci 4(5) 401-405... 

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