# Production of Alternative Fat from Adipose-Derived Stem Cell from Bovine in 3D Culture
## 3 Results
### Adipogenic Differentiation of bADSC on TVP Scaffold for Production of Cultivated Heat  
The production of cultivated meat requires the integration of nutrients, such as fat or protein, with a scaffold to achieve structural similarity to intact tissue. TVP is utilized as a plant-based protein. In this study, an attempt was made to culture bADSCs on TVP coated with a gelatin solution. Given that the perfect adhesion to the TVP is impracticable, assessing the attachment rate of the total cells is imperative. To accomplish this, bADSCs were seeded onto TVP coated with gelatin and incubated for 2 days. TVP-unattached bADSCs would disperse in the culture medium and adhere on the bottom of the cultured flask based on our assumption. Thus, the cells in the supernatant and cultured flask were counted to identify the attachment rate. bADSCs adhered to the TVP were harvested by treating with trypsin-EDTA, and the cell count was determined. (Figure S3). Consequently, the attachment rate ranged approximately from 30% to 44%, with the remaining being unattached under these conditions. Subsequent experiments involved seeding bADSCs to maintain a final cell number of 2 \(\times\) 10\({}^{6}\) within the TVP. bADSC-seeded TVP was incubated for 2 days in growth medium before being transferred to (Figure 3A).  
The differentiation potential of bADSCs was contrasted with that of cell-free TVP. Following the completion of the experimental treatment, the samples were subjected to heating to simulate the processing typically applied to edible tissue. When compared to cell-free TVP, the presence of bADSCs induced changes in both color and texture of TVP. This disparity was further pronounced after heating. Upon heating, TVP with bADSCs exhibited oil release (Figure 3B, red arrow), a phenomenon not observed in the cell-free counterpart. To confirm the adipogenic differentiation of bADSCs, TVP tissue was cryosectioned, and Oil Red O staining was carried out (Figure 3C). As shown in Figure 3C, the presence of lipid droplets within TVP was distinctly detected, confirming the adipogenic differentiation of bADSCs and the production of lipid droplets within the TVP matrix.  
\begin{table}
\begin{tabular}{c c c c c} \hline \hline
**Common Name** & **Fame** & **Abbreviation** & **bAdipocyte** & **Fat Tissue** \\ \hline \multicolumn{5}{c}{**Unsaturated Fatty Acids**} \\ \hline Oleic acid & cis-9-Oleic acid methyl ester & C18:1c & 11\% & 20\% \\ Linoelaidic acid & Methyl linoleicidate & C18:2n6t & 9\% & 3\% \\ Elaidic acid & trans-9-Elaidic acid methyl ester & C18:1t & 8\% & 29\% \\ Eicosatrienoic acid & _cis-8,11,14-Eicosatrienoic acid methyl ester_ & C20:3n6 & 5\% & 1\% \\ Docosahexaenoicenoic acid & _cis-4,7,10,13,16,19-Docosahexaenoicenoic acid methyl ester_ & C22:6n3 & 4\% & N/D \\ Palmitoleic acid & Methyl palmitoelate & C16:1 & 2\% & 4\% \\ Eicosapentaenoic acid & _cis-5,8,11,14,17-Eicosapentaenoic acid methyl ester_ & C20:5n3 & 2\% & N/D \\ Eicosenoic acid & Methyl cis-11-eicosenoate & C20:1n9 & 2\% & 1\% \\ Eicosadinenoic acid & _cis-11,14-Eicosadinenoic acid methyl ester_ & C20:2 & 1\% & N/D \\ Erucic acid & Methyl cruciate & C22:1n9 & 1\% & N/D \\ Docosahexaenoicenoic acid & _cis-13,16-Docosalenoic acid methyl ester_ & C22:2 & 0\% & 0\% \\ Myristoleic acid & _Methyl myristolate_ & C14:1 & N/D & 2\% \\ \hline \hline \end{tabular}
\end{table}
Table 1: _Cont._