Cell Isolation from Tissue (original) (raw)

In the past several decades, cell transplantation has been growing widespread as one of the most promising treatments for different diseases. However, the methods of cell isolation are not optimized at the current time. Therefore, the choice of one technique over another has been often arbitrary and based more on individual experiences rather than on an understanding of why a certain method works and what modifications could lead it to even a better outcome. The goal of a cell isolation procedure is to optimize the yield of functionally viable, isolated cells. Since cell isolation is a complex procedure, many factors affect its outcome. Type of tissue, donor’s body main index and age, cold ischemia during organ preservation, dissociation medium, dissociation enzyme(s), Impurities in any crude enzyme preparation, concentration(s) of enzyme(s), temperature, and digestion times are among the factors that affect the cell isolation outcome. Researchers searching the scientific literature for information on the ideal enzymes and optimal conditions for tissue dissociation are often confronted with conflicting data. Much of the variation stems from the complex and dynamic nature of the extracellular matrix and from the historical use of relatively crude, undefined enzyme preparations for cell isolation applications. This chapter summarizes our knowledge of how these enzymes accomplish the "routine" operations of tissue dissociation and cell harvesting; describes standard lab procedures; offers a logical experimental approach for establishing a cell isolation protocol; and lists many tissue specific references.

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