The biological department of Shanghai Medicilon has extensive experience in the fields of molecular biology, cell biology, in vitro biology and structural biology. From the initial cDNA library construction to drug design, through protein purification, structure determination and analytical determination, we provide a complete set of biological services.
Main content of cell experiment technical service
Stable cell line screening service content and process: 1) Conventional cells: a. Killing curve drawing; b. TransCell Outsourcing Servicefection of plasmids, plating, screening of cell monoclonals with different antibiotic concentrations; c. qPCR or Western Blot to detect the foreign source of stable clones Gene expression level. 2) Special cells: a. According to the cell characteristics, experimental characteristics or experimental needs, design the experimental plan and discuss with the customer to determine the experimental plan; b. Pre-experiment, determine the best transfection reagent by transfecting the plasmid containing the GFP reporter gene, and Optimize the transfection conditions; c. draw the killing curve; d. transfect the plasmid, plate and screen the monoclonal cells with different antibiotic concentration; e. qPCR or Western Blot to detect the expression level of the stable cloned foreign genes.
Primary cell separation experiments: conventional cell culture experiments and construction of cell models for experiments on morphology, gene level and protein (group) level. Service content: cell primary culture, cell subculture, 3D cell culture. Experiment period: depending on the specific experiment.
Content and process of cell transfection experiment service: 1) Plasmid: a. Perform pre-experiment, optimize transfection conditions by transfection of the plasmid containing the reporter gene; b. After the transfection conditions are determined, transfect according to the customer’s protocol; c. qPCR Or Western Blot to detect transfection efficiency. 2) SiRNA: a. Design and synthesize siRNA based on gene sequence; b. Perform pre-experiment, optimize transfection conditions by transfecting negative control siRNA with fluorescent label; c. Verify the silencing efficiency of siRNA by qPCR and Western Blot, Screen the optimal concentration of siRNA.
Cell viability test (MTT method): There are always some cells that die for various reasons in the cell population, and the percentage of viable cells in the total cells is called cell viability. Thiazolan, or MTT for short, can enter the cell through the cell membrane. The amber dehydrogenase in the mitochondria of living cells can reduce the exogenous MTT to water-insoluble blue-purple Formazan crystals and deposit them in the cells. The crystals can be Dimethyl sulfoxide (DMSO) is dissolved, and its light absorption value is measured at 490 nm wavelength using an enzyme-linked immunoassay detector, which can indirectly reflect the number of cells.
Apoptosis experiment: Apoptosis refers to the autonomous and orderly death of cells controlled by genes in order to maintain the stability of the internal environment. Apoptosis is different from cell necrosis. Apoptosis is not a passive process, but an active process. It involves the activation, expression, and regulation of a series of genes; it is not a kind of autoinjury under pathological conditions. Phenomenon, but a process of death actively striving for better adaptation to the living environment.
Flow cytometry: Flow cytometry (Flow Cytometry, FCM) is a rapid quantitative analysis and analysis of cells or other biological particles (such as microspheres, bacteria, small model organisms, etc.) arranged in a single row in the flow Selected technology. The technology is widely used in all aspects from basic research to clinical practice, covering the fields of cell biology, immunology, hematology, oncology, pharmacology, genetics and clinical testing.
Drug-resistant strain induction experiment: In vitro low-concentration gradient increase combined with large-dose discontinuous shock method was used to induce tumor cells to develop drug resistance. Such as: human breast cancer doxorubicin resistant cell line, human breast cancer paclitaxel resistant strain, human lung adenocarcinoma paclitaxel resistant strain, human liver cancer fluorouracil resistant strain, human colon cancer vincristine resistant strain, human oral epithelium Cancer vincristine-resistant strains, human colon cancer paclitaxel-resistant strains, human colon cancer fluorouracil-resistant strains, human ovarian cancer cisplatin-resistant strains, human leukemia doxorubicin-resistant strains, human ovarian cancer paclitaxel-resistant strains, Human gastric cancer cisplatin-resistant strain, human leukemia doxorubicin-resistant strain, human breast cancer doxorubicin-resistant cell strain, human colon cancer vincristine-resistant strain, human liver cancer fluorouracil-resistant strain, human breast cancer anti-estrogen Hormone resistant strains, human breast cancer Tamoxifen resistant strains, mouse hormone-dependent breast cancer cells, etc.
Primary/continuoua cell culture: The growth of cells requires a nutrient environment. The nutrient matrix used to maintain cell growth is called a culture medium. The medium can be divided into liquid medium and solid medium according to its physical state. Isolate the required cells and expanded cells from multicellular organisms and transform the cells in vitro. Observe that the problem of ex vivo cell culture must be solved. Compared with the difficulty of microbial cell culture, it is more difficult to come from multicellular organisms Single cell culture, especially animal cell culture.
Cell viability test: The dehydrogenase in living cells can reduce MTT to a blue-violet product (formazan) that is insoluble in water, and precipitates in the cells, but dead cells do not have this function. Dimethyl sulfoxide (DMSO) can dissolve the blue-purple crystals deposited in the cells. The color of the solution is proportional to the amount of Formazan contained. Finally, the OD value is measured with a microplate reader.
Stable transfection (Stable transfection): Stable transfection or long-lasting transfection is used to establish a cloned cell line in which the transfected target gene is integrated into chromosomal DNA and guides the synthesis of the appropriate amount of target protein. In general (depending on the cell type), the efficiency of forming stable transfected cells is 1 to 2 orders of magnitude lower than the efficiency of transient transfection. The use of selectable genetic markers facilitates the isolation of rare stable transfectants from non-transfected cells.
Transient transfection (Transient transfection): Transient transfection can easily and quickly study and analyze gene function, and the gene expression level is high, but usually only lasts for a few days, mostly used for promoters and other regulatory elements. Generally speaking, the efficiency of transfection of supercoiled plasmid DNA is high. Within 24-72 hours after transfection (depending on various constructions), the analysis results are often used in some reporting systems such as fluorescent protein, β-galactosidase Wait to help detect.
Common laboratory equipment for cell experiment service
Cytology plays a vital role in the life science department. In recent years, cytology has flourished and significant progress has been made. The construction of the cell laboratory is also a necessary laboratory for various cell experiment service companies or universities. The following are some laboratory instruments and equipment:
- CO2 incubator
The CO2 incubator is an essential instrument for cell culture. It provides cells with a comfortable environment for in vitro culture, such as suitable temperature, humidity, pH, O2 concentration, etc. It is mainly used in tissue engineering, in vitro fertilization, neuroscience, cancer Research and other mammalian cell research. Cells are easily infected by various bacteria and microorganisms when cultured in vitro. Therefore, the sterilization and bacteriostatic ability of the CO2 incubator is particularly important.
ESCOCelCulture series CO2 incubator has a stable culture environment, professional sterilization function and unique antibacterial function. Such as 90 ℃, 14Hour can complete the humid heat sterilization process, can effectively kill a variety of microorganisms; Ulpa filter, the chamber achieves air quality of grade 5; stainless steel polished liner, effectively inhibits the adhesion of microorganisms; ESCO patent ISOCIDETM surface antibacterial coating The technology can effectively consciously breed microorganisms on the surface of the cabinet. Experiments have proved that within 24 hours, 99.9% of bacteria on the surface of the cabinet can be eliminated. Reliable performance, simple operation, and perfect design provide the best cultivation conditions for various types of cells, and it is a step closer to helping your scientific dream become a reality!
- Ultra-clean workbench/biological safety cabinet
The cell culture has high requirements on the sterile environment. When the cell is operated, it is generally required to be carried out under 100-level air quality, so the ultra-clean workbench or biological safety cabinet must be equipped.
Singapore ESCO ultra-clean workbench and biological safety cabinet provide you with more safe protection.
- Pure water system
Cells cultured in vitro are particularly sensitive to water quality and require higher purity of water. If the culture water contains some impurities, even if the content is very small, sometimes it will affect the survival and growth of the cells. The endotoxin in the water directly participates in apoptosis. The lower the endotoxin content, the less toxic to the cells and the more beneficial to the growth of the cells. And passaging; too high endotoxin content will directly lead to premature cell aging, and even lead to cell death. Distilled water prepared with a metal distiller may contain certain metal ions and is generally not used as culture water. For the preparation of the culture solution, triple distilled water or ultrapure water prepared by an ultrapure water purification device that has been distilled three times through a quartz glass distiller should be used.
Millipore Corporation of the United States was founded in 1945 and is headquartered in Boston, Massachusetts. It has subsidiaries and offices in more than 30 countries and regions around the world, providing products and technical services for more than 100 countries and regions. The company currently employs more than 5,800 people worldwide. Since the 1960s, Millipore has been committed to the research and development of pure water systems for laboratories, providing ultra-pure water with a daily output of several to thousands of liters for universities and clinical and pharmaceutical laboratories around the world (I Grade), analytical grade water (grade II) and laboratory grade water (grade III) purification systems. Over the years, brands such as Milli-Q, Elix, Super-Q, RiOs, Direct-Q, and Simplicity have been everywhere in various scientific literatures, and have become the authoritative brand in the minds of scientific researchers worldwide.
- Liquid nitrogen tank
In order to preserve the cells, especially the mutant cells or cell lines that are not easy to obtain, the cells should be frozen. The temperature of frozen storage is generally -196℃ of liquid nitrogen, therefore, a liquid nitrogen tank is required. In this environment, normal cells can be kept alive for ten years.
The capacity of the MVE XC series liquid nitrogen tanks ranges from 700 – 5000 tubes and more than 150 to 1,000 cryopreservation tubes. The product is durable, light weight, world-leading manufacturing technology, and has a 5-year vacuum guarantee.
The above is about cell biology experiment, cell experiment outsourcing, cell experiment technology, cell experiment service, etc. The information comes from the official website of Medicilon.