How to Prepare a Single-Cell Suspension from Mouse Spleen Tissue

[@funny11744]

A general question regarding contamination: Is it a routine to spray alcohol absolutely on every object that is inserted in the laminar flow cabinet. But how about the flasks that are transported to the microscope for checking stem cell detaching ? I did not see in any video on KZbin such an operation of spraying the flask before reintroducing it in the laminar flow cabinet !

[@STEMCELLTechnologies]

We follow aseptic technique while in the lab even if it is not shown in the video. If you are worried about contamination, you may spray a paper towel with ethanol and wipe the bottom of the flask. If you have any more questions, feel free to email us at [e-mail].

[@funny11744]

@@STEMCELLTechnologies thank you .

[@LeouIsmaelBanla]

What is the approximate yield of nucleated cells (absolute number) that you obtain when following this method? Thanks so much.

[@STEMCELLTechnologies]

There is variation in the total number of nucleated cells in the mouse spleen. In general, the older the mouse, the lower the splenocyte yield is. However, the most frequently observed splenocyte yield per spleen is 5 x 10^7 to 1.5 x 10^8 cells. If you have any more questions, feel free to email us at [e-mail].

[@User9527_]

Hi, is macrophage also isolated with this kit?

[@STEMCELLTechnologies]

This product is designed to maximize the recovery of dendritic cells from mouse spleen when combined with EasySep™ cell separation technology (e.g.EasySep™ Mouse Pan-DC Enrichment Kit, Catalog #19763 and EasySep™ Mouse Pan-DC Enrichment Kit II, Catalog #19863). For isolation of macrophages, mechanical dissociation is recommended followed by the use of the EasySep™ Mouse F4/80 Positive Selection Kit (Catalog #100-0659). This Tech Tip (www.stemcell.com/technical-resources/methods-library/cell-separation/sample-preparation-and-storage/preparation-of-single-cell-suspensions-from-tissue-or-cell-culture-samples/prepare-single-cell-suspension-from-mouse-spleen.html) might be useful to you. If you have any more questions, feel free to email us at [e-mail].

[@aarushmadhireddy4326]

What type of science is this what would I need to study

[@STEMCELLTechnologies]

Hi, this video outlines an enzymatic method to generate a single-cell suspension from mouse spleen prior to the downstream isolation of dendritic cells.

[@Fredmcz]

I would love to work with you :)

[@STEMCELLTechnologies]

Hi Fred, thanks for reaching out. Please email [e-mail] with the details.