Serum substitute in epithelial cell culture media: nonfat dry milk filtrate

Abstract

A number of milk types and milk fractions were investigated as possible substitutes for serum in cell culture media. A filtrate of reconstituted nonfat dry milk showed promise. Culture fluids containing 5% of the nonfat dry milk filtrate were used to propagate primary and continuous cell cultures, and the cell growth from these cultures was compared with that of cells grown in a serum-containing medium. The nonfat dry milk filtrate-supplemented medium supported the growth of all epithelial cells tested, but two fibroblast-type cultures failed to replicate. Cells grown in the medium containing the milk filtrate grew slowly for 2 to 3 days and then propagated to confluency in 6 to 8 days. Viable cell counts of 9 days were comparable to those of serum-grown cells that had been propagated for 7 days. Cells grown in the milk filtrate could be split 1 to 4 when subcultures were prepared. Cell growth could be stimulated by refeeding on days 2 to 3 or by the addition of 30 microM 2-mercaptoethanol to the growth medium. Virus susceptibility and titer comparisons with poliovirus 1, coxsackievirus B2, echovirus 7, and herpes simplex virus indicated that approximately the same data were obtained when either the nonfat dry milk filtrate-treated or the serum-treated cells were studied. The nonfat dry milk filtrate is inexpensive, is easily prepared, and is a substitute for serum in epithelial cell culture media.

Optimization of the BGM cell line culture and viral assay procedures for monitoring viruses in the environment

“…The initial inoculation of sewage sample number 2 onto BGM cells yielded only coxsackievirus B3, whereas viruses isolated from the L-132 cells were 88% echovirus types 5, 11, and 17, and 12% coxsackievirus B3. However, when a like sample of sewage inoculum from sample number 2 was pretreated with coxsackievirus B3 antiserum and then inoculated onto BGM cells, 39% of the isolates were found to be other coxsackievirus B types, whereas the remaining 61% were echovirus types 4,5,7,11,13,15,17,24, and 25.…”

Thermal Inactivation of Clostridium perfringens Enterotoxin in Buffer and in Chicken Gravy

“…Cell culture assay Stocks of Vero cells purchased from Flow Laboratories, Inc., RockvilIe, MD were passaged by trypsinization and grown as monolayers at 36°C. Growth medium consisted of equal volumes of Eagle's minimum essential medium with Hank's salts and Leibovitz medium with L-glutamine; growth medium was supplemented with 5% newborn calf serum and 5% nonfat dry milk (Fassolitis et al, 1981).…”