Hi all,
I'm looking for ideas to purify 1000 cells from 1.5ml PBS while minimizing cell loss and as clean as possible (not using beads for example). Also standard centrifugation is not an option due to cell loss (although any ideas for special cenrifugation tubes, devices or methods are welcome). Thats a tricky one, I know. Any ideas? Thank you all in advance.
Hi Barak,
There are numerous approaches to purify cells without cell loss and you need to be careful to select the most appropriate one especially when they are only few in numbers. I would like to recommend adhesion based selection methods as they can be rapid and less expensive. For example, the use of adsorbed adhesion proteins in a plate or well format could be very effective to separate the subpopulation of cells expressing the interacting protein. To achieve this, you need to know or predict the cell surface protein interactions specific to your cell type so that you can make the choice of protein for coating the matrix. Hope it helps!
Use a filtration unit. Amicon large MW cut off. Cell strainer from bd or milteyni. Or a track etch filter from Fisher. All of these are fairly inexpensive and very quick. You should pick your filter size based on your cells. I would lean toward a cell strainer from miltenyi, they have many sizes and come enclosed, for ease of use. Even if you don't use a filter, I would recommend you look at miltenyi's website, they're known for cell separation techniques.
It depends on the composition of your cell suspension. Are they adherent cells or suspension cells? Isolated, primer, or breeded? Do you have just the cells in the 1.5 PBS, or is there something else (debris, ecm, dead cells, etc..) present?
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