Isolation, Identification and Cultivation of Mesenchymal Stem Cells Derived from Adipose Tissue of Laying Hens

Today, the intensity and speed of production in the laying hens causes multiple anomalies such as bone and skeletal problems which leads to heavy economic losses. Conducting studies on stem cells obtained from laying hens can improve our knowledge and understanding of the differentiation and molecular mechanisms at cellular level in these birds. For this reason, the aim of this study was to extract mesenchymal stem cells from the fat tissue of laying hens and culture these cells in laboratory conditions.

In this research, mesenchymal stem cells were isolated in a laboratory environment under sterile conditions from fat tissue of laying hens (Hy-Line strain-w36). Then in an standard culture conditions the mentioned cells were separated from other cells by treatment of collagenase enzyme. The obtained cells after several steps of centrifugation and purification were cultured in DMEM-F12 medium containing 10% fetal bovine serum. During 21 days, cell growth process (PDT) and cell colony formation test were performed to evaluate the performance of isolated cells and the ability of cell growth and proliferation.

About 3 to 4 days after culturing the cells, the adherent cells filled 70-80% of the flask. Cells were passaged up to 14 times with normal morphology. To check the speed of cell growth, this test was repeated in passages 2, 4, and 8 in order to investigate the decrease in the ability of cells to grow with increasing age. The average cell growth rate (PDT) for passages 2, 4, and 8 was 37.03, 60.92, and 101.42%, respectively, which indicates the relationship between stem cells' ability to reproduce and self-renew with their age. After 9 days of cell culture, the number of colonies formed from passage 2 and 5 with concentrations of 175, 350, and 520 cells per 2 cm in 6-well plates were counted as 22, 62 and 84, respectively.

The results of this research showed that mesenchymal stem cells extracted from the fat tissue of laying hens have the ability to maintain their basic characteristics during multiple cell divisions and can maintain their morphological structure and specific characteristics such as growth elated function