Fernández-Pernas Pablo, Fafián-Labora Juan, Lesende-Rodriguez Iván, Mateos Jesús, De la Fuente Alexandre, Fuentes Isaac, De Toro Santos Javier, Blanco García Fco, Arufe María C
Grupo de Terapia Celular y Medicina Regenerativa (TCMR-CHUAC), CIBER-BBN/ISCIII, Servicio de Reumatología, Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), SERGAS, Departamento de Medicina, Facultade de Oza, Universidade da Coruña (UDC), As Xubias, 15006, A Coruña, Spain.
Grupo de Proteómica-PBR2-ProteoRed/ISCIII-Servicio de Reumatologia, Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), SERGAS, Universidade da Coruña (UDC), As Xubias, 15006, A Coruña, España.
J Cell Biochem. 2016 Sep;117(9):2097-108. doi: 10.1002/jcb.25515. Epub 2016 Mar 30.
Our group focuses on the study of mesenchymal stem cells (MSCs) from human umbilical cord stroma or Warthońs jelly and their directed differentiation toward chondrocyte-like cells capable of regenerating damaged cartilage when transplanted into an injured joint. This study aimed to determine whether lactogenic hormone prolactin (PRL) or 3, 3', 5-triiodo-L-thyronine (T3), the active thyroid hormone, modulates chondrogenesis in our in vitro model of directed chondrogenic differentiation, and whether Wnt signalling is involved in this modulation. MSCs from human umbilical cord stroma underwent directed differentiation toward chondrocyte-like cells by spheroid formation. The addition of T3 to the chondrogenic medium increased the expression of genes linked to chondrogenesis like collagen type 2, integrin alpha 10 beta 1, and Sox9 measured by quantitative real time polymerase chain reaction (qRT-PCR) analysis. Levels of collagen type 2 and aggrecane analyzed by immunohistochemistry, and staining by Safranin O were increased after 14 days in spheroid culture with T3 compared to those without T3 or only with PRL. B-catenin, Frizzled, and GSK-3β gene expressions were significantly higher in spheroids cultured with chondrogenic medium (CM) plus T3 compared to CM alone after 14 days in culture. The increase of chondrogenic differentiation was inhibited when the cells were treated with T3 plus ML151, an inhibitor of the T3 steroid receptor. This work demonstrates, for first time, that T3 promotes differentiation towards chondrocytes-like cells in our in vitro model, that this differentiation is mediated by steroid receptor co-activator 2 (SRC2) and does not induce hypertrophy. J. Cell. Biochem. 117: 2097-2108, 2016. © 2016 Wiley Periodicals, Inc.
我们的研究团队专注于对来自人脐带基质或华通氏胶的间充质干细胞（MSC）进行研究，以及它们向类软骨细胞的定向分化，这些类软骨细胞在移植到受损关节时能够再生受损软骨。本研究旨在确定催乳激素（PRL）或活性甲状腺激素3,3',5-三碘-L-甲状腺原氨酸（T3）是否在我们的定向软骨分化体外模型中调节软骨生成，以及Wnt信号通路是否参与这种调节。人脐带基质来源的间充质干细胞通过形成球体向类软骨细胞进行定向分化。通过定量实时聚合酶链反应（qRT-PCR）分析，向软骨生成培养基中添加T3可增加与软骨生成相关基因的表达，如II型胶原蛋白、整合素α10β1和Sox9。与未添加T3或仅添加PRL的情况相比，在添加T3的球体培养14天后，通过免疫组织化学分析的II型胶原蛋白和聚集蛋白聚糖水平以及番红O染色均增加。培养14天后，与单独使用软骨生成培养基（CM）相比，在添加CM加T3培养的球体中，β-连环蛋白、卷曲蛋白和GSK-3β基因表达显著更高。当细胞用T3加ML151（一种T3类固醇受体抑制剂）处理时，软骨分化的增加受到抑制。这项工作首次证明，在我们的体外模型中，T3促进向类软骨细胞的分化，这种分化由类固醇受体共激活因子2（SRC2）介导，且不会诱导肥大。《细胞生物化学杂志》117: 2097 - 2108, 2016。© 2016威利期刊公司。
