Lee Joonbum, Kim Dong-Hwan, Suh Yeunsu, Lee Kichoon
Department of Animal Sciences, The Ohio State University, Columbus 43210, USA; The Ohio State University Interdisciplinary Human Nutrition Program, The Ohio State University, Columbus 43210, USA.
Department of Animal Sciences, The Ohio State University, Columbus 43210, USA.
Poult Sci. 2021 May;100(5):101057. doi: 10.1016/j.psj.2021.101057. Epub 2021 Feb 16.
Current research of avian adipogenesis has been dependent on primary preadipocytes culture due to the lack of commercially available immortal preadipocyte cell lines in avian species. In addition to primary stromal vascular cells, primary chicken embryonic fibroblasts (CEF) were suggested as new in vitro models for adipogenesis study, because CEF can be differentiated into adipocytes by a combination of fatty acids and insulin (FI), or all-trans retinoic acid (atRA) alone in the media containing chicken serum (CS). However, there are decreases in differentiation of primary cells due to diverse population of cell types and low adipogenic potential of cells after passages. In the present study, adipogenic differentiation of DF-1 cells, immortal fibroblasts derived from an embryonic chicken, was tested with 4 different medium; 10% fetal bovine serum (FBS), 10% CS, 10% CS with FI, and 10% CS with FI and atRA. Lipid droplets stained with Oil Red O were not shown in DF-1 cells under 10% FBS, appeared with very small sizes under 10% CS, significantly increased under 10% CS with FI, and most significantly accumulated under 10% CS with FI and atRA. In addition, expressions of markers for adipogenesis (Znf423, C/ebpβ, Pparγ, and Fabp4), fatty acid uptake (CD36), triglyceride synthesis (Gpd1, Dgat2), and lipid droplet stabilization (Plin1) were significantly upregulated by supplementation of 10% CS with FI and atRA. Morphological evidence for formation of lipid droplets and dramatic induction of adipogenic marker genes support the adipogenic potential of DF-1 cells, offering DF-1 cells as a new cell model to investigate various research studies involving avian adipogenesis.
由于禽类缺乏可商业化获得的永生化前脂肪细胞系，目前禽类脂肪生成的研究一直依赖于原代前脂肪细胞培养。除了原代基质血管细胞外，原代鸡胚成纤维细胞（CEF）被认为是脂肪生成研究的新体外模型，因为在含有鸡血清（CS）的培养基中，CEF可通过脂肪酸和胰岛素（FI）组合或单独使用全反式维甲酸（atRA）分化为脂肪细胞。然而，由于细胞类型多样以及传代后细胞的脂肪生成潜力较低，原代细胞的分化能力会下降。在本研究中，使用4种不同培养基对源自鸡胚胎的永生化成纤维细胞DF-1进行脂肪生成分化测试；10%胎牛血清（FBS）、10% CS、含FI的10% CS以及含FI和atRA的10% CS。在10% FBS条件下，DF-1细胞中未出现用油红O染色的脂滴；在10% CS条件下，脂滴出现但尺寸非常小；在含FI的10% CS条件下，脂滴显著增加；在含FI和atRA的10% CS条件下，脂滴积累最为显著。此外，通过添加含FI和atRA的10% CS，脂肪生成标志物（Znf423、C/ebpβ、Pparγ和Fabp4）、脂肪酸摄取（CD36）、甘油三酯合成（Gpd1、Dgat2）以及脂滴稳定化（Plin1）的表达均显著上调。脂滴形成的形态学证据和成脂标志物基因的显著诱导支持了DF-1细胞的脂肪生成潜力，为研究涉及禽类脂肪生成的各种研究提供了一个新的细胞模型。
