# Production of Alternative Fat from Adipose-Derived Stem Cell from Bovine in 3D Culture
## 1 Introduction  
Conventional meat production emits large amounts of greenhouse gases, which significantly contribute to global warming [1]. These emissions stem from activities such as feed production and manure management, collectively contributing to 14.5% of global greenhouse gas emissions [2]. Therefore, increasing meat production to meet rising demand is expected to further exacerbate environmental impact. Presently, plant-based alternative meat is widely acknowledged and represents a popular choice among vegan alternatives [3, 4]. Various types of plant-based meat continue to be produced, and textured vegetable protein (TVP) is mainly used as plant-based meat substitute [5, 6, 7, 8]. However, plant-based meat has faced criticism for relying on potent food colorings, textural additives, causing deficiency in essential nutrients and elevated sodium content [9, 10, 11]. Cultivated meat, emerging as a sustainable future food source, is produced by growing animal cells in a controlled environment and is gaining attention as a sustainable and eco-friendly meat for the future [3; 12]. Cell-based meat production is estimated to significantly reduce greenhouse gas emissions by decreasing land use, water consumption, and food-crop demand [13; 14]. This mimics the natural in vivo of cells and resembles cell types arranged in structures as animal tissues, thereby replicating the nutritional/organoleptic profiles of conventional meat. However, less previous research is available in cultured meat on mimicking food-related characteristics, such as sensorial properties and nutritional value, compared to traditional meat studies. Most research in the cultured meat field has concentrated on developing muscle tissue, the primary component of meat products. In contrast, few studies have addressed cultured fat tissue. Lipid composition of the fat tissue in the meat is critical in determining meat quality, affecting flavor, texture, and palatability [15; 16]. A method development for generating cultured fat from animal cells is crucial for mimicking these important food-related attributes. Numerous researchers have sought to induce adipogenesis in animal-derived adult stem cells in vitro to obtain fat for cultured meat. There are various types of adult stem cells in the body. Among them, mesenchymal stem cells (MSCs) have the capability to differentiate into various cell types, including osteoblasts, chondrocytes, and adipocytes [17; 18]. MSCs can be isolated from almost every tissue in the body, such as bone marrow, adipose tissue, trabecular bone, and umbilical cord blood [18; 19; 20]. The global application of MSCs for critical diseases demonstrates their safety and efficacy in the human body. However, challenges in stem cell harvest arise due to the limited quantity of tissues that can be collected. Thus, adipose tissue is emphasized due to its abundant availability and easy access. Adipose-derived stem cells (ADSCs) are MSC subsisted in adipose tissue. Large quantities of ADSCs can be easily harvested from abundant adipose tissue in the body, and these harvested ADSCs have potential to expand rapidly in vitro [21; 22]. ADSCs have excellent capability to generate a multitude of growth factors and differentiate into adipocytes compared to MSCs from other tissues. These distinct characteristics of ADSCs led ADSCs to be the most promising stem cell population, and the utilization of ADSCs in multiple studies enhanced ADSCs value as a cell source for not only the pharmaceutical field but the cultivated meat industry. Recently, the possibility of using ADSCs for the food industry was suggested due to the gradual increase in ADSCs usage as a source for cultivated meat production. The production of cultivated meat necessitates a significant quantity of cells, leading to elevated production costs [22; 23; 24]. In order to achieve a sufficient cellular product, suspension culture employing scaffolds is notable for its ability to support cell growth in a three-dimensional (3D) form [25; 26; 27].