UNSW Cell Biology

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Cells Image

"A picture says a thousand words...."

The above image shows 2 neuronal cells in tissue culture. This neuronal cell line, unlike adult neurons, continue to proliferate and differentiate in tissue culture (in vitro).

The 2 recently divided cells (daughters) show a characteristic "butterfly-like" appearance as the cells migrate in opposite directions away from each other. The final stage of cell division, cytokinesis, can still be seen as the narrow region between the 2 daughter cells.

Each cell also shows cellular features typical of neurons, a round cell body (soma) containing the cell nucleus, and elongating cell processes (neurites). These neurites on neurons in the brain and spinal cord would continue to grow and differentiate into 2 types of processes, axon and dendrites. Axons and dendrites contain different proteins, structures and have different functional roles.

Neurite growth occurs by extending highly motile regions in neurons called growth cones.

A single photograph does not of course show these dynamic and functional changes that are investigated by cell biologists in an attempt to understand how this process occurs and why it goes wrong in disease.

Living Cell Dynamics

This timelapse series shows the motile dynamics of the B35 cell line.

The Neuronal Cell Line

The cell line B35 is a clone, from a single parent cell, generated by chemical-induced mutation in new born rat brains. A number of different clonal rat cell lines were generated by this technique.

Newborn rat brains were dissociated, and each cell line was generated from a single cell (clone).The name of each clone was derived from the cell culture well (in this case B35) in which the original cell (clone) was growing.

These cell lines were first characterised, determining their properties. Cells were then stored by freezing and can be thawed and grown when needed.

References:

1. Schubert D, et al. [See Related Articles] Clonal cell lines from the rat central nervous system. Nature. 1974 May 17;249(454):224-7. No abstract available.
2. Schubert D. [See Related Articles] Induced differentiation of clonal rat nerve and glial cells. Neurobiology. 1974;4(6):376-87. No abstract available.
3. Schubert D, et al. [See Related Articles] Putative neurotransmitters in clonal cell lines. Nature. 1975 Mar 27;254(5498):341-3. No abstract available.

Cell Growth

The cell lines are grown in sterile conditions attached to a surface (a plastic tissue culture plate) covered in liquid growth media.

These tissue culture plates are kept in a heated (37 C) incubator with a gas /air mixture (5% carbon dioxide) designed to maintain physiological pH.

The Image

The tissue culture plates are made from clear plastic and living cells can be viewed by an inverted microscope (light from above, lens views from underneath) using special phase optics.

Without phase optics the cells would be difficult to see. The phase optics give healthy cells (with intact cell membranes) a "halo" effect of white light around the cell.

Rotating lens allow viewing of cells with different magnifications. The "photograph" was taken with a video camera connected to a computer. Scale bar and labels added to the image by a graphics program.