I currently want to dissect a rat and store its organs inside liquid nitrogen for future cells or stem cell extraction, including heart, liver, kidney, bone marrow, tendon, cardiomuscle, etc. I have questions:
1) What kind of media should I use?
2) Can I store them in -86 C Freezer instead of LN2 for long periods of time (few months)?
3) What is the precaution I should notice during the thawing process?
Thanks a lot for your answers,
Long.
Hey Long, look at the two links below that describe the best ways to preserve your tissues for cell culture down the road. Hope this helps.
http://www.andrew.cmu.edu/user/yr25/TaylorPublications/MJTaylor108.pdf
Exp Toxicol Pathol. 1999 May;51(3):229-34.
Cell cultures from cryopreserved renal biopsies and other tissue samples.
Sommer M1, Fünfstück R, Stein G.
Hello
For each type, including a special type of media and the method of preparation of storage
See the attachment
Good luck
Dear Long,
Storage in Liq N2 is preferable but in certain cases people have also tried storage of tissues in -80 freezer and stem cell isolation has been performed lateron. But in case of Liq nitrogen u can obtain more viable cells but I think culturable cells can be obtained even after storage at -80. I have observed the same in our lab that -80 storage doesn,t affect the functional properties of certain stem cells but other stem cells can have a drastic affect,. So preferably u should go for Liq N2. You can opt for -80 for short term storage.. Hope it helps !
Dear Long,
Depends on the particular precursor cells you want to culture. Precursor cells composed of progenitor (lineage-committed cells with defined proliferation potential) and stem cells (lineage-uncommitted cells with unlimited proliferation potential). Progenitor cells composed of unipotent, bipotent, tripotent, multipotent committed cells. Stem cells composed of ectodermal, mesodermal, endodermal, germ layer lineage, pluripotent, and totipotent cells. To isolate Progenitor cells freeze tissues in liquid nitrogen. For ectodermal, mesodermal, endodermal and germ layer lineage stem cells store tissues in -80C. For pluripotent and totipotent stem cells store tissues at 4C.
Dear Long
I agree with Dr Young, anyway here you can find other interesting information:
http://stemcellres.com/content/pdf/scrt397.pdf
Hope it can help
Isolation of the stem cells from fresh tissues is always desirable. However, liquid nitrogen storage of the tissue is an alternative.