3D Spheroid Culture Enhances the Expression of Antifibrotic Factors in Human Adipose-Derived MSCs and Improves Their Therapeutic Effects on Hepatic Fibrosis

Figure 3

Effects of transplanted 3D spheroid-derived MSCs on hepatic fibrosis and liver function. CCl₄ was injected intraperitoneally into adult female C57BL/6 mice to induce hepatic fibrosis. Then, 500 μL of PBS, 1 × 10⁶ of 2D cultured MSCs, or 1 × 10⁶ of 3D spheroid MSCs were injected into the tail vein of hepatic fibrosis model mice. (a) HE and Sirius red staining. Four weeks after MSC transplantation, hepatic tissues were obtained and paraffin-embedded sections were prepared. HE and Sirius red staining were performed to evaluate the hepatic fibrosis (bar = 100 μm). (b-c) Expression levels of fibrosis-associated factors. Four weeks after MSC transplantation, hepatic tissues were obtained and expression levels of fibrosis-associated factors (Collagen I and Collagen III) were determined by Western blotting. GAPDH was the internal control. (d–j) Analysis of serum markers associated with liver function. Four weeks after MSC transplantation, serum levels of albumin (d), ALT (e), TBIL (f), AST (i) were tested using automatic biochemistry analyzer. Serum HA (g) and TGF-β1 (h) levels were determined by ELISA and the double-antibody sandwich chemiluminescence immunoassay, respectively. Serum ALP activity (j) was estimated using commercial kits. compared with normal; compared with PBS and compared with PBS; compared with 2D MSC medium and compared with 2D MSC medium.

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