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Passaging Cells

Overview

Cell lines are frequently used in biomedical experiments, as they allow rapid culture and expansion of cell types for experimental analysis. Cell lines are cultured under similar conditions when compared to freshly-isolated, or primary, cells, but with some basic important differences: (i) cell lines require their own specific growth factor cocktails and (ii) their growth must be more closely monitored than primary cells, as the mutations that allow them to be grown indefinitely also can quickly lead to their overgrowth. Therefore, when a cell line reaches the point of growth in culture where it covers most of the bottom of the culture container, or about a 90% confluency, the cells must be resuspended, washed, used experimentally, frozen for later use, or re-seeded for further expansion in new culture containers.

This video will demonstrate how to use media indicators to determine cell culture health, which reagents and equipment are useful for safely removing adherent cell lines from culture, and various methods for transferring these robustly expanding cells into new cultures will be discussed. Also demonstrated are methods for how to culture feeder cells (important for providing essential growth factors to cell lines) and how to expand large numbers of cell line cultures at once.

Procedure

Passaging, or subculturing, of cells, is a common procedure wherein cells from a given culture are divided, or “split”, into new cultures and fed with fresh media to facilitate further expansion. While the concept itself is relatively simple, in this video we will discuss some of the more important steps for the successful maintenance and expansion of cell lines.

Toxic metabolites accumulate in cell culture media over time. When expanding cells, it is particularly important to

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Tags
Passaging CellsSubculturing CellsCell CultureCell LinesMedia ChangeCell HealthCell ExpansionImmortalized Cell LinesStem Cell LinesHeLa Cell LineCancer CellsSelf renewing CellsMultipotent CellsEmbryonic Stem CellsSuspension Culture

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