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As regarding for protocol of isolation of stem cell from cord...
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Mervat Ahmed Daood Nov 30, 2018
As regarding for protocol of isolation of stem cell from cord tissue by mixed explant and enzymatic method (trypsin ).,Iam work by this protocol yield many many cells and doing sub culture to passage 6 but when do characterization by flow cytometry (BD accuri) Iam found CD 45 N(range from Read more 
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Shan Wang Author Answered Dec 4, 2018
School of Life Sciences, Tsinghua University, China, China,
Dear,
Thanks a lot for your question. I am very pleasure to share my opinions.
From your statement, I think your cells are not purified and most of the cells are blood cells (leucocyte). When you culture these cells, are they attached to the plates and spread well?
When I isolated stem cells from cord, adipose and placenta tissues, I usually seed around 10 million cells in a 10 cm plate. Then I remove at least half of the suspension after 24 hours and add the same volume of fresh medium. After that, I change medium every three days till around 2 week. At this time we can see that there are some colonies formation but nearly no blood cells.
When the colonies are big enough, trypsinized and expand the cells. When the cells are enough, we can do FACS analysis and detect different markers.
Good luck and hopefully it can help you!
Best,
Shan
Thanks a lot for your question. I am very pleasure to share my opinions.
From your statement, I think your cells are not purified and most of the cells are blood cells (leucocyte). When you culture these cells, are they attached to the plates and spread well?
When I isolated stem cells from cord, adipose and placenta tissues, I usually seed around 10 million cells in a 10 cm plate. Then I remove at least half of the suspension after 24 hours and add the same volume of fresh medium. After that, I change medium every three days till around 2 week. At this time we can see that there are some colonies formation but nearly no blood cells.
When the colonies are big enough, trypsinized and expand the cells. When the cells are enough, we can do FACS analysis and detect different markers.
Good luck and hopefully it can help you!
Best,
Shan
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