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

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Ahmadian Farhang; Irani Mehrdad; Mohammadi Sangcheshmeh Abdollah

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Journal Paper

Paper Information

Journal: Research On Animal Production Year:2023 | Volume:14 | Issue:4 Page(s): 62-67

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Information Journal Paper

Title

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

Author(s)

Ahmadian Farhang | Irani Mehrdad | Mohammadi Sangcheshmeh Abdollah | Issue Writer Certificate

Keywords

Bone differentiation

Genistein

Mesenchymal stem cell

Laying hen

Abstract

Background: Today, the intensity and speed of production in Laying hens cause multiple anomalies, such as bone and skeletal problems, which lead to significant economic losses. Conducting studies on stem cells obtained from Laying hens can enhance our understanding of the differentiation and molecular mechanisms at the 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 under laboratory conditions.
Methods: In this research, Mesenchymal stem cells were isolated in a laboratory environment under sterile conditions from the fat tissue of Laying hens (Hy-Line strain W36). Under standard culture conditions, the cells were separated from other cells by treatment with collagenase enzyme. The obtained cells, after several steps of centrifugation and purification, were cultured in DMEM-F12 medium containing 10% fetal bovine serum. Over 21 days, the cell growth process (population doubling time, PDT) and cell colony formation tests were performed to evaluate the performance of the isolated cells and their ability to grow and proliferate.
Results: 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 while maintaining normal morphology. To assess the speed of cell growth, this test was repeated in passages 2, 4, and 8 to investigate the decrease in the ability of cells to grow with increasing passage number. The average cell growth rate (PDT) for passages 2, 4, and 8 was 37.03%, 60.92%, and 101.42%, respectively, indicating a relationship between the stem cells' ability to reproduce and self-renew and their age. After 9 days of cell culture, the number of colonies formed from passages 2 and 5, with concentrations of 175, 350, and 520 cells per 2 cm² in 6-well plates, were counted as 22, 62, and 84, respectively.
Conclusion: The results of this research showed that Mesenchymal stem cells extracted from the fat tissue of Laying hens can maintain their basic characteristics during multiple cell divisions, preserving their morphological structure and specific characteristics, such as growth-related functions.

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