A kind of selective medium for skin fibroblast culture and preparation method thereof

Abstract

The invention provides a selective culture medium for skin fibroblast culture. The selective culture medium comprises a basic culture medium, fetal calf serum, human serum albumin, bovine pituitary extracts, insulin, vitamin C, biotin, insulin-like growth factors and epidermal growth factors which are mixed according to a proportion being 1ml to (0.0125ml to 0.05ml) to (0.005ml to 0.025ml) to (80mug to 423mug) to (7mug to 18mug) to (22mug to 60mug) to (37mug to 214mug) to (1mug to 20mug) to (0.1mug to 1mug). A DMEM culture medium and an F12 culture medium are used for basic culture. The selective culture medium for skin fibroblast culture has the advantages that the selectivity is realized on the fibroblast; the purity of the fibroblast is improved; the sundry cell pollution is avoided; the cell passage number is effectively reduced; the cell proliferation speed is high. The selective culture medium provided by the invention can also reduce the serum consumption; the cost is reduced.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a selective culture medium for skin fibroblast culture and a preparation method thereof. Background technique [0002] Skin fibroblasts are widely distributed in the dermis. Fibroblasts secrete a variety of substances to form the extracellular matrix, thereby forming connective tissue and exerting structural functions on the body. Fibroblasts play an important role in human life, such as guiding the formation of skin in the embryonic stage, participating in the maintenance of skin homeostasis after organ maturation, and promoting the repair of the structure and function of the damaged site. Therefore, culturing autologous skin fibroblasts in vitro has a wide range of applications in the fields of tissue engineering, cosmetology, and medical treatment. [0003] At present, the medium used in the method of culturing fibroblasts in vitro is basically configured by basal medium and 20% f...

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Problems solved by technology

[0004] 1. There is no selectivity for fibroblasts, and it is necessary to perform various complicated operations on primary tissues to obtain relatively pure dermal tissues

For example, the primary tissue is digested with trypsin to separate the epidermis and dermis, and then the dermis is chopped up and digested into single cells with collagenase, and finally cultured in the culture medium. During this operation, the enzyme will cause irreversible damage to the cells. The cell yield is low, and it also has the disadvantages of high cost, long digestion time, complicated operation, etc.

[0005] 2. Due to the lack of fibroblast selectivity, it is often difficult to obtain purified fibroblasts in the first three passages, and multiple passages are required to obtain high-purity cells. However, the more passages will make the primary activity of the cells worse and worse. The curative effect is significantly reduced, and there is a risk of tumorigenesis

Additionally, trypsinization with each passaging can cause severe damage to the cells

[0006] 3. In the subculture operation, in order to ensure the cell viability and proliferation speed, it is impossible to expand the bottle culture in a large number, and often choose 1 bottle to transfer 3 bottles or less, resulting in long culture time, many passages, and poor primary cell activity.

Although the amount of serum used in this medium is small, it is not selective for fibroblasts, and the miscellaneous cells in the medium have a great impact on the growth of fibroblasts

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