## 4 Discussion  
In this study, we evaluated whether the extracted MSCs from human hair follicles have multiple differentiation potentials. Based on the previous studies, the main criteria of cells to be defined as multipotent is their ability to differentiate into various lineages such as chondrogenic, osteogenic, and adipogenic [21; 22]. Therefore, we tested the differentiation capacity of HFMSCs in three different lineages. It was found that HFMSC changed its morphology inside the adipogenic, chondrogenic, and osteogenic differentiation mediums, projecting its multi-differentiation potential, while the differentiation morphology of HFMSCs exhibited a morphological similarity to MSCs [23]. During adipogenic differentiation, it is considered that as long as adipocytes are mature, the oil droplet will be formed in culture MSC [19; 20]. In our study, the mature adipocytes were observed inside the adipogenic medium before day 12 indicated by the droplet formations. Moreover, we also evaluated the ability of HFMSC to differentiate into chondrocyte and osteocyte. Previous studies found that after chondrogenic differentiation, MSCs will gradually condense into a compact structure and can be stained using immunofluorescence [20; 21]. Additionally, mature chondrocytes can also be detected with a glycosaminoglycan marker by Alcian Blue staining [21]. In line with this study, we found HFMSC showed a positive result for Alcian Blue staining. The chondrogenic medium can induce the aggregation of HFMSCs then gradually condense into compact structures. We also discovered that the osteogenic transformation of HFMSCs can be triggered through the osteogenic differentiation medium. Therefore, our results indicated that the isolated HFMSCs have the ability to differentiate into adipocytes, chondrocytes, and osteocytes. It is generally believed that the cell adherence in vitro can be facilitated using various combinations of biomaterials [12; 24; 25]. One approach to this requirement is the establishment of hydrogels via molecular self-assembly of peptides and their derivatives [26; 27]. It has been verified that hydrogels comprise an intrinsic hydrate architecture that imitates the mechanical properties of soft tissue and ECM [28; 29]. Nevertheless, some limitations of the hydrogels such as leak of protein adsorption made this material sometimes poorly promote a direct adherence of the cells [30; 31]. The present study indicated that the derivatives of hydrogel peptide could promote the cells to aggregate into 3D spheres. In addition, we found that the modification of GRGD with F127 could urge the cells to assemble and form a spheroid structure. Similarly, a previous study that used the mixture of F127 with  
Figure 8: Effect of the 3 BE hydrogel with different concentrations on the cell growth evaluated by using CCK assay on day 1, day 4, and day 7 (\(n=3\)): (**a**) Swiss 3T3 and (**b**) HFMSCs. (\({}^{*}p<0.05\)). RGD to culture the adipose-derived stem cells (ASCs) confirmed a higher cell adhesion of ASCs [14]. Supporting this finding, another study found that immobilization of RGD peptide onto macroporous scaffolds enable chondrogenesis of human MSCs by promoting cell adherence [17]. Although, the 3 BE hydrogel could promote the cells to aggregate into 3D spheroids. Some of the spheroids exhibited a slightly irregular shape at this stage (Figure 6). The formation of this feature can be attributed to the influence of nutrient and oxygen gradients along with diffusion kinetics [32]. These are also factors to be considered and overcome when culturing cells in 3D, especially as the uniform size of the cellular aggregate grows using the 3 BE hydrogel in the future. Furthermore, it was found that the utilization of Pluronic F127 as a 3D cell culture substrate is suitable to maintain the growth of HFMSCs. The addition of membrane-stabilizing agents in Pluronic F127 results in maintenance of cell viability over time [13].