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Close Message User Date(yyyy-mm-dd) Use of trypsin inhibitor in primary cell culturing An 2009-03-18 Click here to register. Hi! For my research I am coculturing neurons on top of astrocyte monolayers in serum free medium. However, the plating of primary neurons on top of the astrocytes does not lead to consequently good results. Sometimes the astrocytes start to deform (they become stringy en start to form "holes") and ultimately the cells will die! For the culturing of the neurons, I dissociate hippocampal tissue in a trypsin solution and wash this solution with serum free medium. In our protocols we never use trypsin inhibitors, but I have now some very strong indications that the trypsin might be damaging my cells :s So, I am considering to use trypsin inhibitor. But I would like to know if there might be any cons to do so? (Otherwise I just have to start washing the tissue better and more). Any advice/help/information on this subject will be greatly appreciated!!! Thx!!   Re: Use of trypsin inhibitor in primary cell culturing Grifta 2009-03-23 Click here to register. Try washing the tissue in media with serum in, as this inactivates trypsin. Paul   Re: Use of trypsin inhibitor in primary cell culturing RTE 2009-03-25 Click here to register. Also the lack of serum could be causing problems with the attachment of the primary neurons. A small amount of serum would help this or a specialty serum free medium that contains attachment factors might help.

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mold contamination by jak332 on 2009-07-28 filter out leucine from media? by bristow on 2009-07-08 Re: filter out leucine from media? by lubrol on 2009-07-24 Vendor for INS-1 cell line by achu on 2009-06-19 Difference between High and Low Glucose DMEM by purush_79 on 2009-06-11 HK-2 cells by PFMUR on 2009-06-09 cell culture by choosme on 2009-05-10 symptoms of stress by janaki ram on 2009-05-09 PMA cell activation by melissga on 2009-04-28 tumor spheroids culture by A38 on 2009-04-03 White Whisps in Culture Media by mspada on 2009-04-01 Use of trypsin inhibitor in primary cell culturing by An on 2009-03-18 Re: Use of trypsin inhibitor in primary cell culturing by Grifta on 2009-03-23 Re: Use of trypsin inhibitor in primary cell culturing by RTE on 2009-03-25 Removal of macrophages in murine marrow culture by purush_79 on 2009-03-18 BONE MARROW MAST CELLS CULTURE by masto on 2009-03-17 Food for B16 F10 by LadyJustin on 2009-03-17 CD106 cells in murine marrow by purush_79 on 2009-03-17

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