Tracking Cell Lineages of Single Cells

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Entry by Andrew Capulli, AP225 Fall 2011


"Tracking lineages of single cells in lines using a microfluidic device" Amy C. Rowat, James C. Bird, Jeremy J. Agresti, Oliver J. Rando, and David A. Weitz, Proc. Natl. Acad. Sci. U. S. A. 106(43), 18149–18154 (2009).

Introduction: Motivation

While some cell lines continue to express apparently similar phenotypes over time as they replicate, other cell populations don't replicate at all (or do so very minimally). Take the human body in say, ten years of aging: skin tissue and the cells therein are relatively conserved, meaning the skin you have at the starting point will look and will physically be very similar to your skin in 10 years. Cardiac myocytes, those that make up your heart tissue will also 'look' the same but for a different reason: for the most part, they are the same (these are very general statements but they illustrate the point that follows). Cell expression (protein expression) as a function of time and generation is a somewhat under investigated area. As generally mentioned about tissues, the behaviors of cells in bulk (tissue) can and have been observed but the behaviors of individual cells and cells lines are masked in the three dimensional "ensemble" as the authors call it. In tissues or even just unconstrained clusters of cells (say, in a petri dish of culture flask) the lineages of dividing cells become mixed in the three dimensional space so when staining and investigation into particular phenotypes (protein expression) is investigated, it may be clear which cells are expressing the phenotype but their 'age' or 'generation' is unclear... tracking if the cells are newly spawned is impossible. Therefore, in these simple set-ups it is impossible to track phenotype as it varies down one lineage (conserved genetic makeup) of cell replication (starting with one cell). Here the authors propose to address this question with a novel microfluidic device that allows them to investigate the phenotype of a single cell lineage via separate chambers that house a single cell. The chambers allow for the cell to replicate down the chamber(1 dimensionally) and consequently the authors, after a period of replications, have the consequent generations starting from a single cell assembled in a line where they can then investigate cell phenotype.

Lineage Chamber Device

Much of this paper describes the lineage chamber device as shown below in Figure 1 from the paper. A detailed account is in the paper but I'll now summarize the device for the purpose of discussion later on. Thin chambers (1 budding yeast, Saccharomyces cerevisiae, cell wide) are assembled in parallel (50 chambers in parallel per unit). The chamber can be seen in part A of Figure 1. Cells and media are injected into the device; as the authors describe, once a single cell occupies a chamber, the flow in that chamber is reduced and consequently it is less likely that another cell flows into that chamber. The flow through the device is described by the previous wiki entry that can be found by clicking: fjfj