how to change cell shape? - (Oct/08/2009 )

Does anyone have any ideas or techniques on how to stretch or squeeze a mammalian cell in 2-D? Any and all ideas appreciated! Thanks!

phdgirl on Oct 8 2009, 09:56 AM said:

A 5$ physical posibility,
Imagine a room that get smaller as you advance you there will be a point at which you can advance, but if you were forced to advance you would squeeze as much as you can, till becoming flat. As que have bones we wouldn't become flat but cells don't.
A cell is around 10um two glass plates also separate constantly around 10um. But we want an ever smalling room so we better go a plastic plastic way. To allow an 100um entrance put a piece of tape on each corner on the entrance size. To reach a gradually smalling room exert a gradient of vertical force over the plates so that they will aparently fuse on the exit extreme.
As you place the liquid at the entrance it will get in by itself due to capillarity. To force through the smalling camera use vacuum or electrophoretic force.
This will get you a flat cell.
If you want a needle cell then make pressure in one of the sides of the camera and make the cell progress that way.

Thus, you fist separate your cells by size and flexibility and then make them "needles"

To much force will lyse your cells, to litle wont force them channge shape.

If you want to fix the cells perfuse 4%Paraformaldehyde to the camera.

If you want to inmobilize them perfuse ready to polymerize agarose or polyacrylamide.

Should the cells remain alive?
If so, u should probably mutate some kinetochore or cytoeskeletal genes.

You may also add electroporation in a lipid rich medium if the cells are inbetween two plates you can play with the plates separation to give them the desired shape.

phdgirl on Oct 8 2009, 09:56 AM said:

I've heard that cell micropatterns can define cell shape, making cells spread in specific ways, which may adress your question.

Curr Opin Cell Biol. 2006 Dec;18(6):648-57. Epub 2006 Oct 12.
Cell shape and cell division.
Théry M, Bornens M.

I found out that M bornens is also CSO of a french biotech, called Cytoo. They have some micropatterns product, so if you are interested, you may have a look to their website.

Mechanical response of cells to their environment is becoming an important issue in cell biology. Unidirectional stretching of cells growing on silicone/elastomer (PDMS) membranes has been used in several labs. Application of either cyclic stretch or shear stress to cells is used.

See for example:
Mechanical force mobilizes zyxin from focal adhesions to actin filaments and regulates cytoskeletal reinforcement
Masaaki Yoshigi, Laura M. Hoffman, Christopher C. Jensen, H. Joseph Yost, and Mary C. Beckerle
J. Cell Biol. 2005 171: 209-215.

Depending on the expected effects , and the resolution needed, force can be applied on confluent cells or on individual cells. For individual cells, a possible problem is that cells are not stretched identically, as they have different shapes and sizes with respect to the stretch direction.
Using individual cells growing on adhesive micropatterns printed on the silicone/elastomer membranes is explored in several laboratories. This should considerably simplify the control of force application and also increase the reproducibility of the effects from one cell to the next, since cells are normalized by the adhesive micropatterns.

These techniques are not easily accessible to most academic cell biology labs, as specific equipments are required to make these patterns. One of the reasons for starting CYTOO was to make these approaches more widely accessible.

M Bornens