Combination of PD98059 and TGF-β1 Efficiently Differentiates Human Urine-Derived Stem Cells into Smooth Muscle Cells
| Autor: | Hee-Sook Jun, Donghee Kim, Yongha Hwang, Seon-Heui Cha |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Adult
Male Cell type QH301-705.5 Cellular differentiation Calponin Urine Catalysis Article Inorganic Chemistry Transforming Growth Factor beta1 Western blot Tissue engineering medicine Humans Biology (General) Physical and Theoretical Chemistry QD1-999 Molecular Biology Spectroscopy Flavonoids urine-derived stem cell Muscle Cells pluripotent adult stem cells biology medicine.diagnostic_test diabetes Chemistry Stem Cells Organic Chemistry Cell Differentiation General Medicine Middle Aged Computer Science Applications Cell biology Real-time polymerase chain reaction biology.protein Female differentiation of insulin producing cell Stem cell Adult stem cell |
| Zdroj: | International Journal of Molecular Sciences Volume 22 Issue 19 International Journal of Molecular Sciences, Vol 22, Iss 10532, p 10532 (2021) |
| ISSN: | 1422-0067 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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