Triton, saponin, methanol, etc… are excellent permeabilization reagents, which are useful for fixed cells. Those agents are highly used in immunocytochemistry, when antibodies or other non-permeable cell membrane reagents are going to be used to label cytoplasmic or nuclear molecules. On the other hand, if you want to test the efficacy of an experimental compound as an anti-cancer drug, you need to start with living cells, to later measure its cytotoxic activity. In this case, the experimental drug is cell impermeable and by using liposomes or electroporation, you are going to internalize the experimental compound and keeping initially the cells alive. Subsequently, with the experimental compound inside of the cells, you are going to measure the level of cytotoxicity of your experimental compound. Good luck!

If the compound is impermeable, I would suggest certain structural modifications.Formulations are not

effective in fixing permeability. In any case you can use the following Permeabilization Buffer: 0.5% Triton X-100, 0.2 µg/ml EDTA and 1% BSA in PBS.

How about using some cyclodextrin as they are very well known, cheap, FDA approved and are known to act as a good permeation enhancers.

The compound is water soluble. After treament, how can you find that these compounds remain in the medium? Do these compounds show cytotoxicity in the cancer cells? If the drugs work, I do not think that all the drugs in the medium can absorbed by cells.

Continue with Rob, you can load any liposomes with your drug and used as drug delivery vehicle. Moreover, you can used a commercial one or prepare them by yourself. You will find several publications in the web, regarding how to prepare them. Another, alternative method is electroporation. This method is also effective to internalize molecules into cells; even DNA. On the other hand, if you used Triton, you are going to dissolve the plasma membrane and destroy the cells. Good luck!

use DMSO OR 0.1 N HCL for preparation

Triton, saponin, methanol, etc… are excellent permeabilization reagents, which are useful for fixed cells. Those agents are highly used in immunocytochemistry, when antibodies or other non-permeable cell membrane reagents are going to be used to label cytoplasmic or nuclear molecules. On the other hand, if you want to test the efficacy of an experimental compound as an anti-cancer drug, you need to start with living cells, to later measure its cytotoxic activity. In this case, the experimental drug is cell impermeable and by using liposomes or electroporation, you are going to internalize the experimental compound and keeping initially the cells alive. Subsequently, with the experimental compound inside of the cells, you are going to measure the level of cytotoxicity of your experimental compound. Good luck!

I think first you must be sure that your compound is cytotoxic, and if you can´t get it into cancer cells, how do you know that you really have a promisory anticancer drug?? Anyway, liposomes are the best choice to internalize a water soluble compound

Have you used saponin detergent at less 1% concentation in medium or buffer? Saponin have the capacity to remove some cholesterol molecules from cell membrane.

There are actually two options depending on your "compounds".

1. If your molecule is aprotein, peptide, Ab or DNA etc. you can use the Chariot delivery system from ActiveMotif.

2. If this is a small molecule, the best bet is nanocochelates. You can look here for a n article on cochelation (basically improvements on liposomal delivery)

http://www.drugdeliverytech.com/ME2/dirmod.asp?sid=&nm=&type=Publishing&mod=Publications%3A%3AArticle&mid=8F3A7027421841978F18BE895F87F791&tier=4&id=655CBA9DCE3B4370A8F660D5135A20E3

and it contains some protocols.

The next version of this is nanocochelates that is exemplified by the Bioral kit for nanocochelates available from BioDelivery Sciences International, and is a simple mix and add kit. You can go the company website and request the kit.

here are two papers about them. The authors are in India, so you may be able to contact them directly.

I am having some difficulty finding the Bioral kit anymore, so you may have to do some emailing !!!!

Good luck !

Here is another reference

And....

Thank you to all for your valuable suggestions. Any one can suggest how about microemulsion (Oil in water) instead of liposomes? Is it helpfull for DNA kind of a molecule for its delivery?