During isolation of hair cells what was the protease enzyme used to get the individual cells. Is there any method available to identify hair cells using antibody?
Hi there,
The following should help in some way:
http://www.worthington-biochem.com/tissuedissociation/Skin.html
I think some fundamental questions have to arise in answering this considering the tissue type and vendour sources, All in all, I have laid hands on trypsin-EDTA and it is the most I come across by ThermoFisher comp. I would advise you read many protocols online and highlight to link the source of the most suitable to your study. Well done.
@dominique correct me if I am wrong. The article says it's a hair cell culture so it is taken from fully grown cochlea. Separation of inner hair cells and out hair cells itself a cumbersome process. And the article had no information of how the cell which was cultured is only hair cells. Normally this kind of neuron eptheilial cell are very sensitive to trypsin edta. I hope some other protease enzyme which work in the presence of calcium only can help in tissue disintegration. The author had never cited a paper where a filling grown hair cell is been isolated and made as continuous cell line. More over this a neuronal cell which not redifferentiate after some passage. I believe so many information in this article is not clear and it seems to you hide from sunlight just by keeping your palm in front the face.
@kamoru A thanks for your answer. Generally while doing FACS, we won't prefer trypsin instead accutase is more likely used. In this case it's hair cell and that too it has calcium dependent potassium channel in it and do thing this EDTA and Trypsin combination will hamper the hair cell isolation?. To solve this Mistry only author should reveal how exactly it was possible to him.
Dear author I am very much interested in this project can you please clarify my question. Project is really very interesting
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