Method validation of multipotential differentiation of human induced pluripotent stem cells in vitro

FENG Dingli; ZHUO Lidan; LU Di; LI Hong; GUO Hongyan

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Medical Journal of the Chinese People Armed Police Forces ›› 2019, Vol. 30 ›› Issue (4) : 303-307.

ORIGINAL ARTICLES

Method validation of multipotential differentiation of human induced pluripotent stem cells in vitro

FENG Dingli^1,2, ZHUO Lidan¹, LU Di³, LI Hong⁴, GUO Hongyan¹

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Abstract

Objective To study the multi-directional differentiation ability of induced pluripotent stem cells (iPSC). Methods iPSCs were induced into Mesenchymal stem cells (MSCs)-like cells by the formation of Embryonic body EBs. The morphology of cells was observed under inverted microscope. Flow cytometry was performed. Expression of surface markers of iPSC-derived MSCs (iPS-MSCs), and further induction of iPSC-MSCs into osteoblasts, chondrocytes; or direct EB inoculation, which induces them into nerve cells. The multi-directional differentiation ability of iPSC was verified by the above method. Results After induction, iPSC-MSC gradually grew into a long spindle shape; CD29 and CD105 were positive in iPSC-MSCs, while CD34 and CD45 were negative; alkaline phosphatase and toluidine staining showed iPSC- MSCs have the ability to form osteoblasts and cartilage; iPSCs can also be induced into neurons directly from the EB stage. Conclusions Through the above methods, iPSC can be successfully induced into osteoblasts, cartilage and neuroblasts, which provides a technical basis for further research and application of iPSC.

Key words

induced pluripotent stem cells / mesenchymal stem cells / cell differentiation

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FENG Dingli, ZHUO Lidan, LU Di, LI Hong, GUO Hongyan. Method validation of multipotential differentiation of human induced pluripotent stem cells in vitro[J]. Medical Journal of the Chinese People Armed Police Forces. 2019, 30(4): 303-307

References

[1] Takahashi K,Yamanaka S.Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors[J].Cell,2006,126(4):663-676.
[2] Hubbard J J, Sullivan S K, Mills J A, et al. Efficient iPS cell generation from blood using episomes and HDAC inhibitors[J]. J Vis Exp, 2014 (92):e52009.
[3] Trokovic R, Weltner J, Nishimura K, et al. Advanced feeder-free generation of induced pluripotent stem cells directly from blood cells[J]. Stem Cells Transl Med, 2014, 3(12):1402-1409.
[4] 肖枫林, 李清刚, 傅博, 等. 干细胞构建类肾体的研究进展[J]. 中国医药, 2016, 11(12): 1882-1886.
[5] Engle S J, Puppala D. Integrating human pluripotent stem cells into drug development[J]. Cell Stem Cell, 2013, 12(6): 669-677.
[6] Sampaziotis F, De Brito M C, Geti I, et al. Directed differentiation of human induced pluripotent stem cells into functional cholangiocyte-like cells[J]. Nature Protocols, 2017, 12(4):814-827.
[7] Burridge P W. Li Y F, Matsa E, et al. Human induced pluripotent stem cell-derived cardiomyocytes recapitulate the predilection of breast cancer patients to doxorubicin-inducedcardiotoxicity[J]. Nat Med, 2016, 22(5): 547-556.
[8] Takasato M, Er P X, Chiu H S. et al. Kidney organoids from human iPS cells contain multiplelineages and model human nephrogenesis[J]. Nature, 2015, 536(7615): 564-568.
[9] McCracken K W, Catá E M, Crawford C M, et al. Modelling human development and disease in pluripotent stem-cell-derived gastric organoids[J]. Nature, 2014, 516(7531):400-404.
[10] Watson C L, Mahe M M, Múnera J, et al. An in vivo model of human small intestine using pluripotent stem cells[J]. Nat Med, 2014, 20(11):1310-1314.
[11] Dye B R, Hill D R, Ferguson M A H, et al. In vitro generation of human pluripotent stem cell derived lung organoids[J]. eLife, 2015, 4: e05098.
[12] Schwartz M P, Hou Z, Propson N E, et al. Human pluripotent stem cell-derived neural constructs for predicting neural toxicity [Neuroscience][J]. Proc Natl Acad Sci U S A, 2015, 112(40):12516-12521.
[13] Hariharan K, Kurtz A, Schmidtott K M. Assembling kidney tissues from cells: the long road from organoids to organs[J]. Front in Cell Dev Biol, 2015, 3(3 Pt 2): 1-9.
[14] Mungenast A E, Siegert S, Tsai L H. Modeling Alzheimer's disease with human induced pluripotent stem (iPS) cells[J]. Mol Cell Neurosci, 2016, 73:13-31.
[15] Lee C T, Bendriem R M, Wu W W, et al. 3D brain organoids derived from pluripotent stem cells: promising experimental models for brain development and neurodegenerative disorders[J]. J Biomed Sci, 2017, 24(1):59.
[16] Nishimura T, Hatoya S, Kanegi R, et al. Feeder-independent canine induced pluripotent stem cells maintained under serum-free conditions[J]. Mol Reprod Dev, 2017, 84(4):329.
[17] Ikeda M, Imaizumi M, Yoshie S, et al. Implantation of induced pluripotent stem cell-derived tracheal epithelial Cells[J]. Ann Otol Rhinol Laryngol, 2017, 126(7):517-524.
[18] Dirja B T, Yoshie S, Ikeda M, et al. Potential of laryngeal muscle regeneration using induced pluripotent stem cell-derived skeletal muscle cells[J]. Acta Oto-Laryngologica, 2016, 136(4):6.
[19] Zhu H, Kimura T, Swami S, et al. Pluripotent stem cells as a source of osteoblasts for bone tissue regeneration[J]. Biomaterials, 2019,196:31-45.
[20] Hashimoto A, Naito A T, Lee J K, et al. Generation of induced pluripotent stem cells from patients with duchenne muscular dystrophy and their induction to cardiomyocytes[J]. Int Heart J, 2016, 57(1):112-117.
[21] Nam Y, Rim Y A, Ju J H. Chondrogenic pellet formation from cord blood-derived induced pluripotent stem cells[J]. J Visual Exp Jove, 2017, 2017(124):58-63.
[22] Chen Y S, Pelekanos R A, Ellis R L, et al. Small molecule mesengenic induction of human induced pluripotent stem cells to generate mesenchymal stem/stromal cells[J]. Stem Cells Transl Med, 2012, 1(2):83-95.
[23] Zheng Y L, Sun Y P, Zhang H, et al. Mesenchymal stem cells obtained from synovial fluid mesenchymal stem cell-derived induced pluripotent stem cells on a matrigel coating exhibited enhanced proliferation and differentiation potential[J]. PLoS One, 2015, 10(12):e0144226.