Conclusions

A number of assays are available to the preservation biologist. Fluorescence probes have now proven to be the best choice for accomplishing viability and functional assays given their abundance, versatility, and high sensitivity. New instruments, such as Molecular Devices, FLIPR system designed for high throughput analysis, may allow preservation biologists to look at multiple fluorescence probes in a near-real time format so that changes in cell physiology (e.g., mitochondrial potential, intracellular calcium, etc.) can be followed over the course of several days subsequent to return to normothermic temperatures. Furthermore, it is important that multiple assays be chosen to assess viability and function as long as they are not measuring the same physiological parameter. This problem can be avoided if assays are chosen from Tiers 1, 2, and 3 as described in this article. Finally, there needs to be a focus on the Tier 4 assays that are more diagnostic in nature so that improved preservation protocols and solutions can be developed.

Jf-ML

Jf-ML

12 hr C3A-averaged fold change graph

12 hr C3A-averaged fold change graph

FIGURE 6.11 Ciphergen SELDI-TOF proteinchip protein profile of cells cryopreserved in different cryo-preservation solutions. SELDI-TOF analysis of cytoplasmic proteins from C3A cells (human hepatoma cell line) cryopreserved in either cell culture media supplemented with 5% DMSO (M5) or CryoStor CS5 (CS5). The panel of 11 profiles (top) is a representative sample of raw SELDI-TOF data, whereas the graph below plots protein abundance against molecular weight. Red bars indicate proteins that increase in abundance in cells cryopreserved in CryoStor CS5; whereas the blue bars represent proteins that increase in abundance in cells cryopreserved in cell culture media supplemented with 5% DMSO.

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FIGURE 6.11 Ciphergen SELDI-TOF proteinchip protein profile of cells cryopreserved in different cryo-preservation solutions. SELDI-TOF analysis of cytoplasmic proteins from C3A cells (human hepatoma cell line) cryopreserved in either cell culture media supplemented with 5% DMSO (M5) or CryoStor CS5 (CS5). The panel of 11 profiles (top) is a representative sample of raw SELDI-TOF data, whereas the graph below plots protein abundance against molecular weight. Red bars indicate proteins that increase in abundance in cells cryopreserved in CryoStor CS5; whereas the blue bars represent proteins that increase in abundance in cells cryopreserved in cell culture media supplemented with 5% DMSO.

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