Combination of PD98059 and TGF-β1 Efficiently Differentiates Human Urine-Derived Stem Cells into Smooth Muscle Cells
- Authors
- Hwang, Yongha; Cha, Seon-Heui; Kim, Donghee; Jun, Hee-Sook
- Issue Date
- Oct-2021
- Publisher
- MDPI
- Keywords
- Diabetes; Differentiation of insulin producing cell; Pluripotent adult stem cells; Urine-derived stem cell
- Citation
- International Journal of Molecular Sciences, v.22, no.19
- Journal Title
- International Journal of Molecular Sciences
- Volume
- 22
- Number
- 19
- URI
- https://scholarworks.bwise.kr/gachon/handle/2020.sw.gachon/82939
- DOI
- 10.3390/ijms221910532
- ISSN
- 1661-6596
- Abstract
- Pluripotent adult stem cells have potential applications in cell therapy and tissue engineering. Urine-derived stem cells (UDSCs) differentiate into various cell types. Here, we attempted to differentiate human UDSCs (hUDSCs) into smooth muscle cells (SMCs) using transforming growth factor-beta 1 (TGF-β1) and/or PD98059, an extracellular signal-regulated kinase (ERK) inhibitor. Both quantitative polymerase chain reaction (qPCR) and Western blot analysis showed that the expression of messenger ribonucleic acid (mRNA) and proteins for alpha-smooth muscle actin (α-SMA), calponin (CNN1), and smooth muscle myosin heavy chain (SM-MHC), which are specific markers for SMCs, increased on day 9 after differentiation and again on day 14. The differentiated cells from human UDSCs (hUDSCs) with a combination of TGF-β1 and PD98059 showed the highest expression of SMC marker proteins. Immunocytochemical staining performed to assess the molecular expression revealed CNN and α-SMA colocalizing in the cytoplasm. The cells that differentiated from hUDSCs with a combination of TGF-β1 and PD98059 showed the strongest expression for CNN1, α-SMA, and SM-MHC. Functional testing of the differentiated cells revealed a stronger contractile capacity for the cells differentiated with a combination of PD98059 and TGF-β1 than those differentiated with a single factor. These results suggest the combination of PD98059 and TGF-β1 to be a more effective differentiation method and that differentiated SMCs could be used for restoring the functions of the sphincter muscle or bladder. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
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