Primary cells of human thymic epithelium

Primary cells of human thymic epithelium

Training Information:

Wrapped condition: collagen I in mouse tail（2-5Mg/cm2）

culture mediumcontainFBSGrowth additivesPenicillinTheStreptomycinwait

Fluid change frequency per2-3Change the fluid once every day

Growth characteristics: Wall adhesion

Cell morphology epithelial cell like

Passage characteristics can be passed down1-2generation

digestive juice0.25%

Cultivation conditions gas phase: air,95%；CO2，5%

Human thymic epithelial cells isolated from thymic tissue; The thymus is an important lymphatic organ in the body, and its function is closely related to immunityTThe place where cells differentiate, develop, and mature can also secrete thymic hormones and hormone substances, which are organs with endocrine skills. Thymic epithelial cells and thymic cells are important components of the thymic microenvironment. In addition, thymic epithelial cells form the three-dimensional structure of thymic cells at different developmental stages. According to their different positions in the thymus, they can be divided into cortical thymic epithelial cells and medullary thymic epithelial cells. The development and maturation of thymocytes are achieved through interactions during the migration of thymic cortical and medullary epithelial cells. In addition, thymocytes develop into mature cells capable of recognizing and tolerating their own major histocompatibility complexes and self antigens through positive selection mediated by cortical thymic epithelial cells and negative selection mediated by medullary thymic epithelial cellsTLymphocytes. Thymic epithelial cells are the main components of the thymic microenvironment, with diverse morphology, distribution, and function. The expression of surface antigens is highly heterogeneous, forming different types of complexes with thymocytes, and some thymic epithelial cells are arranged to form vesicular structures. Electron microscopy observation can divide thymic epithelial cells into3-6Type, using monoclonal antibodies against thymic epithelial cells for fluorescence staining, thymic epithelial cells can be classified into9Types.

Method Introduction:

The human thymic epithelial cells isolated in the company laboratory were treated with protease-The collagenase mixed digestion method combined with differential adhesion method was prepared by culturing and screening with epithelial cell specific culture medium, with a total cell count of approximately5×10?cells/Bottle.

Quality inspection:

Human thymic epithelial cells isolated from the company laboratoryCytokeratin-18Immunofluorescence identification, purity can reach90%Above, and not containingHIV-1TheHBVTheHCVMycoplasma, bacteria, yeast, fungi, etc.

① Organizational block cultivation method

Tissue block culture is a commonly used, simple, and highly successful primary culture method. The basic method is to inoculate the cut small tissue clusters into a culture bottle (or dish), and the bottle wall can be pre coated with a thin layer of collagen to facilitate the adhesion of the tissue blocks to the bottle wall, allowing surrounding cells to grow outward along the bottle wall.

② Digestive cultivation method

③ Suspension cell culture method

For cells that grow in suspension, such as leukemia cells, lymphocytes, bone marrow cells, cancer cells and immune cells in pleural and ascites, digestion is not required. They can be isolated by low-speed centrifugation and cultured directly, or cultured by inoculating after lymphocyte stratification.

④ Organ culture

Organ culture refers to the direct cultivation of organs or tissue blocks obtained from donors under specific environmental conditions outside the body without tissue separation. Organ culture can maintain the relative integrity of organ tissues and can be used to focus on observing the connections, arrangements, and interactions between cells, as well as the biological regulatory effects of local environments.

After receiving the cells, please proceed with the following steps for the customer. (The number of passages for this cell is limited to about 2. It is recommended that the customer arrange subsequent experiments as soon as they receive the cells.)

1. Remove the cell vial, disinfect it with 75% alcohol, remove the sealing film, and place it in a 37 ℃, 5% CO2 incubator for 6-8 hours to stabilize the cell state.

2. When the cells reach 80% confluence, prepare for subculture.

3. Cell passage:

1) Suck out the culture medium from the cell vial and wash the cells once with PBS.

2) Add about 1mL of 0.125% digestive solution to the culture bottle and digest at 37 ℃ for about 3 minutes; Under the microscope, after the cells shrink and become round, discard the digestion solution and add culture medium to terminate digestion.

3) Gently blow and mix with a straw, inoculate and passage in appropriate ratios such as 1:2 or 1:3, then add fresh culture medium to 5mL and culture in a 37 ℃ 5% CO2 cell incubator.

4) After the cells adhere to the wall, culture and observe, and replace with fresh culture medium every 2-3 days.

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