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An Optimized Protocol for the Generation of iPSC Lines from Lymphoblastoid Cell Lines Using the Non-integrative Sendai Virus
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DOI：10.1002/cpz1.70342.png)
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Abstract
En 中文
The choice of cell source for generating induced pluripotent stem cells (iPSCs) is influenced by several factors, including accessibility, reprogramming efficiency, and the invasiveness of the cell collection procedure. Traditional sources such as skin fibroblasts or peripheral blood mononuclear cells often require invasive sampling, which can pose logistical and ethical challenges, especially in large-scale or retrospective studies. In contrast, many biobanks have established collections of easily accessible lymphoblastoid cell lines (LCLs). These LCL repositories constitute an underutilized yet invaluable resource for iPSC research, owing to their widespread availability and existing genetic and phenotypic annotations. Here, we present a robust and reproducible protocol utilizing Sendai virus vectors to reprogram LCLs into iPSCs. Although LCLs are known to exhibit reduced reprogramming efficiency compared to other somatic cell types, our approach optimizes conditions to improve yield and consistency, providing a reliable pathway for generating high-quality iPSCs from archived samples. This methodology enables the reproducible generation of high-quality iPSCs from archived LCLs, eliminating the need for new biospecimen collection. By providing a detailed and optimized protocol, this work lowers technical barriers and facilitates broader adoption of LCL-derived iPSCs across the research community. Ultimately, it expands access to iPSC technology for large-scale, retrospective, and collaborative studies in human development and disease. Current Protocols published by Wiley Periodicals LLC.Basic Protocol 1: Reprogramming lymphoblastoid cell lines using Sendai virusSupport Protocol 1: Pluripotency confirmation by spontaneous trilineage differentiation with TaqMan (TM) hPSC Scorecard (TM) panel analysis.
Keywords:
Cellular reprogramming
induced pluripotent stem cell (iPSC)
Lymphoblastoid cell line (LCL)
Sendai virus
pluripotency