Human induced pluripotent stem cells (hiPSCs) have core properties of unlimited self-renewal and differentiation potential and\nhave emerged as exciting cell sources for applications in regenerative medicine, drug discovery, understanding of development,\nand disease etiology. Key among numerous criteria to assess pluripotency includes the in vivo teratoma assay that has been widely\nproposed as a standard functional assay to demonstrate the pluripotency of hiPSCs. Yet, the lack of reliability across methodologies,\nlack of definitive clinical significance, and associated expenses bring into question use of the teratoma assay as the ââ?¬Å?gold standardââ?¬Â\nfor determining pluripotency.We propose use of the in vitro embryoid body (EB) assay as an important alternative to the teratoma\nassay. This paper summarizes the methodologies for creating EBs from hiPSCs and the subsequent analyses to assess pluripotency\nand proposes its use as a cost-effective, controlled, and reproducible approach that can easily be adopted to determine pluripotency\nof generated hiPSCs.
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