Abstract

   The aim of the present study was to investigate the expression of
   related genes during the differentiation process of baNCSCs into
   adipocytes using transcriptomics technique, thereby clarifying the
   potential mechanism underlying baNCSCs differentiation into adipocytes
   and providing insights into lipid metabolism and regulation of lipid
   deposition in ruminants. Transcriptomic analysis was conducted on the
   adipocytes of baNCSCs on days 0 (CON0), 3 (DIF3), and 9 (DIF9) of
   differentiation. The results showed that in the early stage of
   adipocyte differentiation of baNCSCs, differentially expressed genes
   (DEGs) are mainly involved in metabolic pathways such as chromosome
   modification, cell cycle progression, and regulation of stem cell
   pluripotency. In the middle and late stages of differentiation, DEGs
   are mainly involved in metabolic pathways such as changes in cell
   morphology and synthesis of fatty acids and triglycerides. Predicting
   the top 10 core hub genes (CHGs) in the protein–protein interaction
   (PPI) network that regulate various differentiation stages of
   adipocytes reveals that ERBB2, EGFR, and MYC are upregulated during the
   early differentiation stage. In contrast, ITGB1, KRAS, CCND1, ACTB,
   VEGFA, MET, and HRAS are downregulated. During the middle and late
   stages of differentiation, the expressions of TP53, CASP3, STAT3,
   CTNNB1, JUN, EGFR, and MYC are upregulated, while IGF1R, PTEN, and HRAS
   are downregulated. In conclusion, the primary enrichment pathways of
   DEGs vary at distinct stages of adipocyte induction and differentiation
   in baNCSCs.

1. Introduction

   Adipocyte differentiation is a complex process, involving pluripotent
   stem cells or precursor cells being regulated by a series of
   transcriptional regulators under the influence of internal and external
   factors, and eventually transforming into mature lipid-filled
   adipocytes [[32]1]. The core of this process lies in the timing
   expression of key genes such as peroxisome proliferator-activated
   receptor γ (PPARγ), fatty acid binding protein 4 (FABP4),
   CCAAT/enhancer binding protein α (C/EBPα), and adipose triacylglyceride
   lipase (ATGL), which regulates metabolic processes such as glycerol
   metabolism, fatty acid oxidation, and carbohydrate transformation in a
   network pattern [[33]2,[34]3,[35]4]. The formation of adipocytes is
   closely related to the quality of animal meat [[36]5]; therefore,
   studying the mechanism of adipocyte differentiation is of great
   economic significance for livestock and poultry breeding.

   Neural crest stem cells (NCSCs) are progenitor cells with pluripotent
   differentiation potential, which can not only differentiate into
   neurons and glial cells but also into mesoderm and ectoderm derivatives
   and serve as transitional structures during early vertebrate embryonic
   development [[37]6,[38]7]. This multidirectional differentiation
   potential makes NCSCs an important candidate cell source for
   regenerative medicine and tissue engineering [[39]8]. Adipogenesis is
   one of the differentiation pathways of NCSCs [[40]9]. In particular,
   adipose-derived neural crest stem cells (AD-NCSCs) have been selected
   due to their neural crest origin, limited adipocyte differentiation,
   and greater experimental specificity, making them more suitable for
   targeted studies on fat formation than stromal vascular fraction cells
   or intramuscular adipose-derived stem cells [[41]9,[42]10]. Adipocyte
   differentiation involves various signaling pathways and regulatory
   factors, leading to changes in lipid profiles both on the cell surface
   and internally [[43]11,[44]12]. By tracking and comparing the
   alterations in cellular biomarkers at different stages of NCSCs
   differentiation, the biological processes (BPs) and stages of
   adipogenesis can be identified.

   Jinnan cattle is one of the five major yellow cattle breeds, primarily
   distributed in the southern region of Shanxi Province, China. This
   breed serves not only as traditional draft cattle but also plays a
   significant role as an important beef cattle breed [[45]13]. Currently,
   the breeding efforts for high-quality Jinnan cattle predominantly
   emphasize meat production. The quality of the meat, characterized by
   attributes such as marbling, tenderness, and flavor, has a direct
   impact on the market price of beef and consumer preferences [[46]14].