In situ growth, freezing and testing of cultured cells
View Patent ↗The present invention provides methods and compositions for the in situ growth, freezing and testing of cultured cells. In particular, the present invention provides methods and compositions for the long-term preservation of cells in ready-to-use formats for testing. In addition, the present invention provides rapid and easy to use means to diagnose viral and other infections. Furthermore, the present invention provides easy to use means to grow and store cells in situ for testing methods. Indeed, the present invention makes viral, chlamydial and other diagnostic methods accessible to small laboratories, including those without cell culture capabilities.
1. A method for the detection of at least one toxin in a sample, comprising the steps of:
a) providing:
i) a cell culture comprising human foreskin fibroblast cells, wherein said cell culture has been frozen and thawed in situ on a substrate, wherein the in situ frozen and thawed cells are suitable for detecting at least one toxin, and
ii) a sample suspected of containing at least one toxin;
b) adding said sample to said cell culture on said substrate without incubating said cell culture after thawing, to produce an inoculated culture;
c) incubating said inoculated culture to produce an intoxicated culture; and
d) observing said intoxicated culture for the presence of cytopathic effect on said cells of said cell culture, thereby detecting said at least one toxin.
2. The method of claim 1 , wherein said substrate is selected from the group consisting of glass and plastic.
3. The method of claim 2 , wherein said glass is a glass coverslip.
4. The method of claim 2 , wherein said plastic substrate is the well of a multiwell plate.
5. The method of claim 1 , wherein said toxin is a Clostridium toxin.
6. The method of claim 1 , wherein said toxin is a Clostridium difficile toxin.