Application of serum bottle
The serum bottle is made of low-extraction borosilicate glass, and the colorless clear bottle meets the requirements of USP Type I and ASTM E 438 Type I Standard Class A. It can be autoclaved at 121℃ for 20min, and can be divided into 125ml, 250ml, 500ml, 1L, 2L according to the volume.
Serum bottle is a container for storing serum, which is generally made of PET material by injection stretch blowing process. The medium is the nutrient required for cell growth. According to its source, it is divided into synthetic medium and natural medium, both of which can be stored in serum bottles.
Serum bottle
Natural medium:
The most commonly used natural medium is serum, basically calf serum. Serum contains a variety of cell growth factors, adhesion-promoting factors and their multi-active substances. Combined with synthetic medium, cells can proliferate and grow smoothly. Serum needs to be stored in special serum bottles at -5°C to -20°C.
Synthetic medium:
Synthetic medium is strictly formulated according to the type and quantity of substances required by cells. There are many types and known components, which facilitates the control of experimental conditions. Contains carbohydrates, amino acids, lipids, inorganic salts, vitamins, trace elements and cell growth factors. However, compared with the natural medium, some natural unknown components cannot be replaced by known chemical components. Therefore, the basic synthetic medium used in cell culture must also add a certain amount of natural medium components to overcome the synthetic culture. Insufficient base, the most common practice is to add calf serum.
The shape of the serum bottle is square, easy to grasp, resistant to most acid and alkali corrosion, the embrittlement temperature is -70 °C, and it still has a certain toughness at -30 °C. It is a good packaging container, whether it is natural medium or synthetic culture. can be stored in serum bottles.
For the isolation, culture and identification of human umbilical cord blood endothelial progenitor cells and the establishment of acellular vascular scaffolds by improved freeze-thaw method
To explore the feasibility and clinical application value of endothelial progenitor cells isolated and cultured from human umbilical cord blood.
Methods: The umbilical cord blood of the fetuses undergoing full-term caesarean section in the Department of Obstetrics and Gynecology, the First Affiliated Hospital of Bengbu Medical College was collected, placed in a serum bottle containing 50U/ml heparin under aseptic conditions, stored in a 4°C ice box, and transported to cells In the culture room, the umbilical cord blood mononuclear cells were obtained by density gradient centrifugation. The obtained mononuclear cells were divided into two parts, and fetal calf serum M199 medium (FCS-M199) and autologous serum M199 medium (AS-M199) were added. And they were seeded in culture dishes with and without human fibronectin (HFN) (marked as: F(0), F(1), A(0), A(1)) respectively. In vitro induction, differentiation and expansion were performed.
The morphological changes of adherent cells during the whole induction and differentiation process were observed, and they were identified from different angles combined with related techniques such as immunohistochemistry, immunofluorescence and flow cytometry.
Results: After 12 days of culture, cord blood mononuclear cells began to adhere. After 3d, the spindle shape began to appear, and then the number of adherent cells gradually increased and gradually became spindle-shaped. When cultured for up to one week, a typical colony formed with spindle cells around and round cells in the center. When the cells were cultured for 14 days, the interconnected cell clusters gradually connected and formed a network-like structure. At the same time, with the continuous increase of the culture time, the long spindle cells also began to gradually shorten, and showed a typical paving stone-like change.
The adherent cells were cultured for one week and it was observed that the number of cells in the medium with human fibronectin was significantly higher than that without human fibronectin, and there was no significant change in the number of cells in the two different mediums.
