A method of cultivation of alveolar epithelial stem cells has been developed
Directed differentiation of stem cells is achieved by using specific differentiation factors, including growth factors.
American scientists from Yale University received type I pneumocytes (alveolar epithelial cells of the lungs) using a special system to recreate the conditions of the microenvironment in the respiratory system.
Scientists used a special rotating machine that creates a unique interface between the liquid and air. Type II pneumocytes obtained from induced pluripotent stem cells (iPSC), or isolated from lung tissue, were placed in such environment, and differentiated into type I pneumocytes.
IPSCs obtained from the alveolar cells (pneumocytes) type II did not differ in molecular phenotype from mature pneumocytes type II, isolated from native lungs. The efficiency of differentiation of these cells into pneumocytes type I were compared in rotating bioreactor and in the traditional cultivation system using small molecules.
Differentiation dynamics were assessed by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), flow cytometry and immunocytochemistry.
Cells cultured in a rotating bioreactor had higher levels of expression of markers specific to type I pneumocytes: aquaporin-5 (AQ5), caveolin-1 and T1α.
Overall, the study showed that the rotating bioreactor creates interface between air and liquid, effectively stimulates the differentiation of type II pneumocytes, including those derived from the iPSCs, into the alveolar cells type I.
Thus, the proposed method allows to obtain a large number of alveolar epithelium cells for tissue engineering and regenerative therapy, as well as for studies of the effectiveness and safety of medicines.