Does anyone know where I can find water-soluble testosterone suitable for cell culture? Sigma has discontinued theirs and I have been unsuccessful in finding another. Can water-insoluble testosterone be used for cell culture? Thank you.
Hello, for our cell culture (Androgen receptor transactivation assay) we use Dihydrotestosterone (DHT, 5α-androstan-17β-ol-3-one) instead. DHT is dissolved in DMSO (10 mM stock), and added to the culture medium (final conc. 200 pM) before the assay.
You can use testosterone or dihydrotestosterone for activation of the androgen receptor. You can use the organic form. No need to use water soluble testosterone.
You can dissolve it in 100% ethanol and dissolve it stepwise in cell culture medium.
You can also use 100% DMSO and do the same. Sollibilty remains good to levels of 5x10-11 M and lower.
Serum can have a positive influence on the solubility of very lipofiel compounds.
Be aware the serum should be treated with activated dextran coated charcoal to extract the steroids from the bovine serum in advance to prevent cross contamination..
Hello, I dissolve the water insoluble stimuli in ethanol and make a stock solution 100x or 1000x concentrated (related to the highest concentration used in my experiments) in PBS.
Agree with the others that you can dissolve the testosterone in DMSO or ethanol. Just a word of caution. The solvents may be cytotoxic at high concentrations. Do a solvent toxicity assay on your cultures first (easily done using an XTT or similar assay) to determine levels of solvent that will not affect the cell. Cell lines differ in their sensitivities for the solvents and it is always better to check this out first.
Most chemical companies like Sigma have synthetic steroids. Since they are a restricted drug under FDA regulations, background checks and inventory keeping are required. However, we have been informed that as soon as the steroids are placed in DMSO, they are no longer classified as restricted drugs. Lot of paperwork for us, but DMSO solves a lot of problems.
Dear Ebony,
Try Tocris. www.Tocris.com
Another possibility in Steraloids. www. steraloids.com
Bye
Willem
Seems I was misinformed by my EHS colleagues. Dissolving in DMSO does NOT make it exempt from US DEA regulations. We've just had to fill out a whole bunch of state and Federal forms to obtain a license to buy and use testosterone. I believe some modified testosterone compounds do not require a license, so you really need to check this out before you order.
Hi All,
I am having a related though not identical issue. I can disolve the tesosterone in 100% EtOH. For my experiments I need to dilute the Ethanol to under 0.1% preferably in HEPES buffer (I have tried a range of buffers). Once it is in 50%EtOH:50% HEPES buffer the tesosterone comes right out of solution. I have tried the same with DMSO with the exact same results.
The testosterone powder I have is T1500 from Sigma. Does anyone have experience with this problem / does anyone have a supplier and code for the testostereone/DHT they are using that may work as an alternative in this particular case / does anyone know if it remains in solution in particular buffer&media over others / or if anyone has heard of a water soluble testosterone since the post was created?
Thanks so much in advance.
All the best,
Kate
My first questions would be what concentration stock solution are you making? and what is your final concentration?
Hello, you can disolve the testosterone in ethanol and then make a dilution greater than 1/10000 for ethanol in the final solution.
See this link https://www.sigmaaldrich.com/catalog/product/sigma/t6147?lang=en®ion=IQ
What would be the storage conditions for 5α-androstan-17β-ol-3-one after dissolving it in ethanol or DMSO?
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