How to make human epidermal keratinocytes to form cell culture?(2)
1. When can we carry out the transmission of ES/iPS cells?
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Right now, the most popular medicine in medicine is stem cell therapy.
This is because stem cells have the potential to proliferate and differentiate, which can be differentiated into the cells that people need under certain conditions, which the medical community calls "universal cells".
So, when is it possible to carry out the transmission of ES/iPS cells?
Cell generation can be carried out when one of the following situations occurs:
(1) MEF trophoblast cell growth over two weeks.
(2) Cloning is too big, or too dense.
There is more and more differentiation in the cultivation of ES clones.
2. during mononuclear cells (monocytes) separation of experimental points is what?
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Mononuclear cells, monocytes) is a kind of important in the body without grain white blood cells, derived from bone marrow hematopoietic stem cells, mature mononuclear cells were released into the blood, constitute about 5% to 5% of peripheral blood leukocytes.
Mononuclear cells (monocytes) are the main points of separation experiment?
(1) In order to obtain better results, the individual nuclear cells of peripheral blood should be used immediately after separation.
(2) All the operation should be performed in 18 ~ 20 ℃.
(3) Carefully cover various Percoll layers to avoid damaging its interface.
(4) Washing and separating the cells 3 times to remove residual Percoll layers and platelets.
(5) If the prepared cells are still not pure, they can be used to remove the remaining NK, T and B lymphocytes with anti-cd2, anti-cd3, anti-cd19 antibody molecules.
3. What should we pay attention to when the hippocampus neurons of the newborn Wistar rats are cultured?
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The hippocampus, a limbic system of the brain, is an ancient cortical structure that plays a crucial role in learning, recent memory, emotional and visceral function regulation.
Neurons in the hippocampus of the brain are highly concentrated, sensitive to a variety of pathogenic factors, and are one of the areas where nervous system diseases such as epilepsy and alzheimer's disease are prone to attack.
In vitro, the primary culture of successful hippocampal neurons can provide a good experimental platform for the study of pathogenesis of nervous system diseases and screening of neuroprotective drugs.
What should we pay attention to when the hippocampus neurons of the newborn Wistar rats are cultured?
(1) Split the organization quickly and minimize operating time.
The blood cells can be separated when centrifuged with Optiprep media.
(2) The liquid that is used to separate tissue cannot be partial to base, which can obviously affect cell status.
(3) After digestion, the tissue releases DNA, which causes the tissue to be thick and can't be blown away. DNAase can effectively alleviate this phenomenon. If found in the above DNAase amount, it is still thick enough to increase the amount of DNAase.
The thick material can not be screened out when sifting, the board will soon stick to the bottom of the board, the washing board cannot be washed away, and there are no cells in the surrounding area, which will seriously affect the success of the experiment.
(4) Cell density is at least 1 x 106/ hole (6 orifice plate) when the plate is grown, and the thin cells cannot grow.
(5) After 6 hours, the cells should be washed in time. After a long time, the debris is not easy to wash off and affect cell growth.
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