苏淮猪背最长肌FAPs细胞体外成脂能力及其基因表达模式的研究

doi:10.11843/j.issn.0366-6964.2023.10.012

Abstract

Abstract: This study aimed to obtain higher purity of fibro/adipogenic progenitors (FAPs) from longissimus dorsi muscle of Suhuai pig, and evaluate the alteration of the adipogenic capacity and gene expression patterns during in vitro culture. The longissimus dorsi muscle of one-day Suhuai boar was digested to obtain suspended mixed cells. In order to determine the optimal differential adhesion time for FAPs cell, the purity of cell isolated from 4 different adhesion time were evaluated by immunofluorescence using an antibody against platelet-derived growth factor receptor α (PDGFRα). Then, the adipogenic differentiation ability of different passage of FAPs cells (P1, P3 and P5) cultured in vitro was compared, and the RNA-Seq was used to identify the differentially expressed genes (DEGs), KEGG pathways and GO functional classification in FAPs cells with different passages.The results showed that the purity of FAPs cells isolated from longissimus dorsi muscle of Suhuai pig by differential adherence for 30 min was much higher than other conditions. Evaluation of the adipogenic capacity of FAPs cells with different passages showed that the adipogenic ability of FAPs cells decreased significantly with the increase of passages in vitro. RNA-Seq results showed that the gene expression patterns of P3 and P5 generation FAPs cells were similar. Compared with P1 to P3 or P1 to P5 generation cells respectively, a total of 2 336 common DEGs was identified, including 1 102 up-regulated DEGs and 1 234 down-regulated DEGs. KEGG and GO analysis showed that the common up-regulated DEGs were mainly enriched in PI3K-AKT-FoxO, PI3K-AKT-HIF-1 and cGMP-PKG signaling pathways etc., and the common down-regulated DEGs were mainly enriched in DNA replication and repair signaling pathways etc. Among them, the genes of IRS1, FOXO3, CCNG2, EGFR and FGF1, which could inhibit cell proliferation and adipogenic differentiation were up-regulated, while the genes of CSF1R and SIPA1L2, which could promote cell proliferation were down-regulated. Compared P3 with P5 generation cells, the up-regulated DEGs were mainly enriched in NOD-like, MAPK and non-canonical Wnt signaling pathways etc.; The down-regulated DEGs were mainly enriched in VEGF, PPAR, Notch and IL-17 signaling pathways etc. Among them, TNFAIP3, AREG, FGF1 and FGF9 genes, which could inhibit adipogenic differentiation were up-regulated, and PPARγ, C/EBPα and LCN2 genes, which could promote adipogenic differentiation were down-regulated.These results showed that porcine FAPs cells with higher purity can be obtained by differential adherence for 30 min. The adipogenic capacity of porcine FAPs is continuously decreased during in vitro culture, and the gene expression patterns also undergo significant changes during in vitro culture. The major change for in vitro cultured FAPs is the reduction of gene expression related to cell proliferation and adipogenic differentiation. This study could provide a reference for developing a medium formula, which could maintain the adipogenic capacity of porcine FAPs cells during in vitro culture.

Key words: pig, FAPs cell, in vitro, adipogenic capacity, gene expression

CLC Number:

S828.2

JI Zhengyu, NI Mengru, ZHANG Zhaobo, ZHAO Gan, HUANG Zan, LI Pinghua, HUANG Ruihua, HOU Liming. Research on Adipogenic Capacity and Gene Expression Pattern of in Vitro FAPs Cells Derived from Longissimus Dorsi Muscle of Suhuai Pig[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4126-4142.

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Research on Adipogenic Capacity and Gene Expression Pattern of in Vitro FAPs Cells Derived from Longissimus Dorsi Muscle of Suhuai Pig

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