The basic formula of serum-free medium: the basic ingredients are the basic medium and the added components.
Most of the cells used for biopharmaceutical and vaccine production are adherent growth or facultative adherent growth when cultured in vitro; when they are grown in serum-free and protein-free medium, due to the lack of various adhesions in serum Adhesion factors such as fibronectin, laminin, collagen, vitreous epithelin, cells often grow in suspension.
The added components include the following categories of substances:
(1) Adhesion promoting substance: Many cells must adhere to grow in order to grow. In this case, serum-free medium must be added with adhesion promoting and expansion factors, generally extracellular matrix, such as fibronectin, laminin, etc. . They are also important mitogens and differentiation factors that maintain normal cell function, and play an important role in the reproduction and differentiation of many cells. Fibronectin mainly promotes adherence and differentiation from mesoderm cells, including fibroblasts, sarcoma cells, granulocytes, renal epithelial cells, adrenal cortical cells, CHO cells, myoblasts, etc.
(2) Growth promoting factors and hormones: add different growth factors for different cells. Hormones are also important substances that stimulate cell growth and maintain cell function. Some hormones are essential for many cells, such as insulin.
(3) Enzyme inhibitors: To culture adherent cells, it needs to be digested and passaged with trypsin. It is necessary to contain enzyme inhibitors in serum-free medium to stop the enzyme digestion and protect the cells. The most commonly used is soybean pancreatin; inhibitor.
(4) Binding proteins and transport proteins: common such as transferrin and bovine serum albumin. The addition of bovine serum albumin is relatively large, which can increase the viscosity of the medium and protect the cells from mechanical damage. Many serum-free media grown in rotary culture contains bovine serum albumin.
(5) Trace elements: Selenium is the most common.
Suggest:
1. Serum-free culture is usually, in the first 24 hours, myocardial cells beat normally, then the pulse rate slows down, 72 hours after the cells need to give external stimulation (such as gently touching the culture dish) and then beat!
2. No neurotransmitter such as norepinephrine, nor transferrin added-so can't answer (according to the actual needs of your experiment)
3. No components are added to the serum-free medium-DMEM medium + double antibody (double antibody may not be added)
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