how to start cord blood culture - (Nov/28/2008 )

i m very new in this area of research and want to isolate stem cells from cord blood of human.so how much cord blood should i take to isolate mononuclear cells for plating in one 6 cm culture plate.kindly guide me

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first of all, what kind of stem cell you trying to isolate?
from CB, you can get CD34+ cell, but could isolate MSC as well
but I am just assume that you are collecting CD34+ cells. (similar protocol for MSC too)

in that case, you will start from density gradient separation to isolate mononuclear cells
most people use ficol solution to do so, (if necessary dont forget to dilute CB before ficol)
then,
you can stain the MNC with appropriate antibody and use magnetic sorting/sorter to isolate CD34+ cells
Thats' it.

also, you asked about how much CB you need. I cannot tell exact amount you need
because some CB is very rich in stem cell, others are not. Also final cell number is depends on your isolation technique as well.

hope this helps

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