Hydrocortisone (Cat. No. HY-N0583) is prepared at 10 mM per mL in DMSO. I would like to dilute it to 0.5 mg/mL for use in stem cell culture. How should I proceed? What diluent should I use, such as 1× PBS? Thank you for your advice!
10 mM corresponds to 3.62 mg/mL. DMSO is toxic to cells, and normal cells can typically tolerate a DMSO concentration of 0.1%. It is recommended to dissolve the product in DMSO to prepare a stock solution, which is then diluted 1,000-fold with cell culture medium to obtain the working solution.
The stock solution you received is 10 mM (3.62 mg/mL). Diluting this 1,000-fold will not meet the requirement for your working solution of 0.5 mg/mL (equivalent to 1.37 mM).
You may freeze-dry 1 mL of the 10 mM product to recover the powder and re-dissolve it in DMSO to prepare a stock solution of 500 mg/mL (or purchase the powder form and dissolve it). Our experimental data indicate that 200 mg/mL is achievable, but the solubility at 500 mg/mL is uncertain. You may test a small amount to verify if it can be fully dissolved.
Most literature reports IC50 values in the micromolar (μM) range.
The answer to this question is provided by MedChemExpress Technical Support.
Hi Marine,
I am not an expert in stem cell culture - however your concentration of 0,5 mg/mL (I assume you mean final concentration) sound rather high. This corresponds to a final concentration of 1,4 mM (see Qiu Weiwei math).
However, to my knowledge normal concentrations for cell culture are around 1 µM ( see https://cdn.stemcell.com/media/files/manual/10000005591-MAN_00.pdf) and Qiu Weiwei comment regarding IC50
or 0,004 - 5 µg/mL (see https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/379/357/h0135use.pdf?srsltid=AfmBOooxivIbhoH6wFt3s1VMZwOV5KAZSD76Q4h04o6Tm8HRdeUp8s-7).
I don't know your exact application but maybe you should double check the final concentration of hydrocortisone you need.
If you would need a 1 µM you could very well dilute your hydrocortisone 1000-fold.
However, according to the handling instruction of MCE the final DMSO concentration should be below 0,5% (see https://file.medchemexpress.com/HandlingInstruction/Compound%20Handling%20Instructions.pdf)
However, 0,1% might be safer
Additional information you will find here (https://file.medchemexpress.com/batch_PDF/HY-N0583/Hydrocortisone-DataSheet-MedChemExpress.pdf)
I would also recommend to check the references MCE is providing to find more information about concentrations that are used in cell culture.
KR Dörthe
Hi Marine. We always use hydrocortisone in our MCE (mouse epithelial cell) growth media and the final concentration we use is 0.4pg/ml. So our stock is diluted in H2O at 100ug/ml and then 4.2ml is added to 1L of media. I don't know if there is a reason you need to use such a high concentration. Even if you needed a much higher concentration, it should dissolve in media at 100ug/ml and you wouldn't need DMSO at all.
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