Authors
Creative Bioarray
Summary
For decades, cell lines have played a critical role in scientific developments. In most cases, researchers just got data generated from cell lines. However, due to some weaknesses of cell lines, scientists become increasingly cautious about these generated results. But now the game has changed! Primary cells now are believed to be a more biologically relevant tool than cell lines for studying human and animal biology. And we design this primary cell culture guide aimed at showing new investigators the basic principles of primary cell and some practical culture skills.
Introduction
Primary cells are cells isolated directly from human or animal tissue by using enzymatic or mechanical methods. After isolation, they are placed in an artificial environment and established for growth. According to the genus (by species or tissues) from which they are isolated, they can be categorized in epithelial cells, fibroblasts, keratinocytes,melanocytes, endothelial cells, muscle cells, hematopoietic and mesenchymal stem cells.
Primary cells differ from cell lines in many ways.
Primary cells have a finite lifespan. After a certain period of time in culture, they will die. And the amount of time primary cells survives in culture varies depending on the cell type. On the contrary, an immortalized or continuous cell line has acquired the ability to proliferate indefinitely.
Primary cell cultures are harsh, requiring optimized growth conditions, including the addition of specific cytokines and growth factors for propagation in serum-free or low-serum growth media, which is absolutely different from immortalized cell lines.
Primary cells retain the natural feature of the tissue where they had been isolated. And due to this character, they are physiologically normal and thus theoretically provide unaltered, “natural” experimental results. Immortalized cell lines, on the other hand, are often transformed with other inducible modifications, which may alter the outcome of experiments.
Procedure
Generally, primary cells retain normal morphology, cellular function, growth characteristics, cellular markers, signaling and genetic integrity when propagated in culture. That's why they are commonly used as in vitro tools for pre-clinical and investigative biological research. Meanwhile, primary cells from different species may be used which can highlight potential differences between humans and preclinical test species. And before in vivo studies, mouse or rat primary cells can be used to refine doses and reduce the number of animals required for preclinical toxicology for cost-saving. In addition, the use of primary cells containing none mutations and chromosomal abnormalities has served as the best representative indicators of normal cell phenotype and progression of the early-stage disease. They also can be used to determine the accuracy of extrapolating human data from an animal model.
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