THE CHARACTERISTICS OF RAT EMBRYONIC FIBROBLASTS (REFS) AND RAT BONE MARROW DERIVED MESENCHYMAL STEM CELLS (RAT-BMMSCS) FROM THE WISTAR RAT: A COMPARATIVE STUDY
Keywords:Antigen-specific, Characteristics, Differentiate, Rat-BMMSCs, REFs
Objective: Rat embryonic fibroblasts (REFs) and rat bone marrow-derived mesenchymal stem cells (rat-BMMSCs) an be used as in vitro models for a variety of studies, including for degenerative diseases such as arterial ischemia, tissue engineering and development of induced pluripotent stem cells (iPSCs). Therefore, the further developments of the use of these two cells of great importance.
Methods: The experiments were performed with Wistar rat, those with 15-17 d gestation (aged 32 w) as a REFs source and those aged 12 w as a BMMSCs source. Dulbecco's modified eagle medium (DMEM) was used for both cell cultures but with different media supplements. Proliferation ability was determined for both by calculating population doubling time (PDT). Characterization was performed by differentiation testing into osteocyte, chondrocyte and adipocyte cells by staining with Alizarin Red, Alcian Blue and Oil Red O and by an investigation of specific antigen characteristics using flow cytometry with positive CD90 and CD29 and negative CD34 markers.
Results: Morphologically, the REFs and rat-BMMSCs had the same fibroblasts like shape. PDT was higher for the REFs than the BMMSCs (p<0.05), and both could differentiate into osteocytes, chondrocytes and adipocyte. The characteristics of the positive markers (CD29 and CD90) were higher in rat-BMMSCs than in REFs.
Conclusion: In this study demonstrated that the explant method for REFs isolation and flushing method for rat-BMMSC isolation are both effective. It also showed that rat-BMMSC grow faster than REFs, and that both cells have the same differentiation ability as rat-BMMSCs but with different specific surface antigen characteristics.
Cong S, Cao G, Liu D. Effects of different feeder layers on culture of bovine embryonic stem cell-like cells in vitro. Cytotechnology 2014;66:995–1005.
Xi G, Hu P, Qu C, Qiu S, Tong C, Ying Q. Induced neural stem cells generated from rat fibroblasts. Genomic Proteomics Bioinfomatics 2013;11:312–9.
Ye J, Hong J, Ye F. Reprogramming rat embryonic fibroblasts into induced pluripotent stem cells using transposon vectors and their chondrogenic differentiation in vitro. Mol Med Rep 2015;11:989-94.
Llames S, Garcia Perez E, Meana A, Larcher F, del Rio M. Feeder layer cell actions and applications. Tissue Eng 2015;21:345-53.
Zschemisch N, Eisenblätter R, Rudolph C, Glage S, Dorsch M. Immortalized tumor-derived rat fibroblasts as feeder cells facilitate the cultivation of male embryonic stem cells from the rat strain WKY/Ztm. Springer Plus 2014;3:588-99.
Song K, Huang M, Shi Q, Du T, Cao Y. Cultivation and identification of rat bone marrow-derived mesenchymal stem cells. Mol Med Rep 2014;10:755-760.
Ichim TE, O’Heeron P, Kesari S. Fibroblasts as a practical alternative to mesenchymal stem cells. J Transl Med 2018;16:212.
Gala K, Anna Burdzinska A, Idziak M, Makula J, Paczek L. Characterization of bone-marrow-derived rat mesenchymal stem cells depending on donor age. Cell Biol Int 2011;35:1055–62.
Kisselbach L, Merges M, Bossie A, Boyd A. CD90 expression on human primary cells and elimination of contaminating fibroblasts from cell cultures. Cytotechnology, 2009;59:31–44.