Culture Process and Applications of Human Alveolar Epithelial Type II Cells

Type II alveolar epithelial cells (AT2) play a key role in maintaining the integrity and function of the alveoli. Until recently, the role of damage to the epithelial repair mechanism after injury in the pathogenesis of idiopathic pulmonary fibrosis has been confirmed, and AT2 cells have been transferred to the focus of attention. Therefore, it has become necessary to use primary human AT2 cells instead of cell lines for in vitro experiments.

Alveolar epithelial cells include two types of special epithelial cells, namely alveolar epithelial type I cells (AT1), which cover about 93% of the alveolar surface area and conduct gas exchange through them, and type II alveolar epithelial cells (AT2) that constitute alveolar cells 60%, and is a producer of different surface-active proteins. AT2 cells play a key role in maintaining the integrity and function of the alveoli and serve as AT1 cell progenitors. The damage to the lungs of substances such as cigarette smoke, viruses, and environmental particles is mainly aimed at alveolar epithelium, emphasizing its role in tissue homeostasis. Until recently, the role of damage repair mechanism in the onset of idiopathic pulmonary fibrosis after the injury has been confirmed, and AT2 cells have been the focus of attention. Therefore, it has become desirable to use primary human AT2 cells instead of cell lines (such as A549 epithelial cells) for in vitro experiments. Some research groups have developed methods to isolate human AT2 cells, all applying tissue digestion (using trypsin or elastase) and continuous filtration in their protocol.

Human primary alveolar epithelial cells from Creative Bioarray were isolated from normal human lung tissue. Human primary alveolar epithelial cells were grown in T25 tissue culture flasks pre-coated with gelatin-coating solution for 2 minutes, then incubated in Creative Bioarray culture complete growth medium for 3-7 days, and then expanded. Before transportation, the cells of the third passage were separated from the flask and immediately frozen and stored in the vial. Each vial contains at least 0.5x106 cells per milliliter. Our method for isolating primary epithelial cells is developed based on the combination of existing methods and our proprietary methods. Incubate the cells with EpCAM-1 (CD326) antibody, and then apply magnetic beads pre-coated with the secondary antibody.

Using antibodies, E-Cadherin or ZO-1 rabbit polyclonal antibody, the expression of markers in human primary alveolar epithelial cells from Creative Bioarray was tested by immunofluorescence staining. All cells were negative for mycoplasma, bacteria, yeast and fungi. Not all donors and/or cell batches have detected HIV-1, hepatitis B and hepatitis C. Upon request, a certificate of analysis will be provided for each cell batch purchased. Under the cell culture conditions specified by Creative Bioarray, cells can be expanded in a split ratio of 1:2 or 1:3 for 2-4 generations. Repeated freezing and thawing of cells is not recommended.

Human primary alveolar epithelial cells can be used for cell-cell interaction, adhesion, PCR, Western blotting, immunoprecipitation, immunofluorescence flow cytometry, or analysis of cell-derived derivatives for required research applications. Creative Biorray ships frozen cells on dry ice. Upon receipt, immediately transfer the frozen cells to liquid nitrogen (-180°C) until they are ready for experimentation. Live cell transportation is also available on request. The primary battery must never be kept at -20°C. Human primary alveolar epithelial cells from Creative Bioarray are used for laboratory research purposes only. Our products may be used in humans, in vitro diagnostic procedures, or therapeutic procedures without authorization.

https://www.creative-bioarray.com/Human-Alveolar-Epithelial-Cells-CSC-C9223J-item-41627.htm