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Induced Pluripotent Stem Cells01:13

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Stem cells are undifferentiated cells that divide and produce different types of cells. Ordinarily, cells that have differentiated into a specific cell type are post-mitotic—that is, they no longer divide. However, scientists have found a way to reprogram these mature cells so that they “de-differentiate” and return to an unspecialized, proliferative state. These cells are also pluripotent like embryonic stem cells—able to produce all cell types—and are therefore...
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Stem cells are undifferentiated cells that divide and produce different cell types. Ordinarily, cells that have differentiated into a specific cell type are terminally differentiated; however, scientists have found a way to reprogram these mature cells so that they dedifferentiate and return to an unspecialized, proliferative state. These cells are pluripotent like embryonic stem cells—able to produce all cell types—and are called induced pluripotent stem cells (iPSCs).
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Reprogramming alters the gene expression in somatic cells, transforming them into induced pluripotent stem (iPS) cells over several generations. Scientists can reprogram cells by introducing genes for four transcription factors—Oct4, Sox2, Klf4, and c-Myc (OSKM) by viral or non-viral methods. These factors are also known as Yamanaka factors after Shinya Yamanaka, who first generated iPS cells using mouse skin cells. Yamanaka was awarded the Nobel Prize in Physiology or Medicine in 2012...
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Updated: Jan 14, 2026

RNA-based Reprogramming of Human Primary Fibroblasts into Induced Pluripotent Stem Cells
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Construction of CD163-knockout induced pluripotent stem cells using reprogramming technology.

Yaya Zhao1, Xiaolong Wu1, Liming Yuan1

  • 1Shaanxi Stem Cell Engineering and Technology Research Center/College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, China.

Theriogenology
|October 25, 2025
PubMed
Summary

Researchers generated a CD163-deficient porcine induced pluripotent stem cell line (CD163-KO iPSC) from knockout pigs. This novel cell line serves as a vital in vitro model for studying Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) infection mechanisms.

Keywords:
CD163PRRSVPluripotencyReprogrammingiPSC

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Area of Science:

  • * Molecular Biology and Virology
  • * Stem Cell Biology and Regenerative Medicine
  • * Animal Biotechnology

Background:

  • * Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) infection relies on the CD163 receptor protein for viral entry.
  • * A robust in vitro cellular model is crucial for investigating PRRSV-host interactions and developing antiviral strategies.
  • * Previous work established CD163-knockout transgenic pigs, providing a foundation for this study.

Purpose of the Study:

  • * To generate a CD163-deficient porcine induced pluripotent stem cell (iPSC) line.
  • * To establish a reliable in vitro model for studying PRRSV infection mechanisms and host-pathogen interactions.
  • * To provide a platform for developing antiviral strategies against PRRSV.

Main Methods:

  • * Isolation of porcine auricular marginal fibroblasts (PAMFs) from CD163-knockout pigs.
  • * Reprogramming using porcine-derived OSKM factors (OCT4, SOX2, KLF4, c-MYC) via a Tet-On lentiviral system and LBCSV protocol.
  • * Characterization of generated iPSCs using alkaline phosphatase staining, qPCR, semi-quantitative PCR, immunofluorescence, Western blot, and RNA-seq analysis.

Main Results:

  • * Successful generation of a CD163-deficient porcine induced pluripotent stem cell line (CD163-KO iPSC).
  • * Confirmed pluripotency through alkaline phosphatase staining and upregulation of key markers (NANOG, SALL4) at mRNA and protein levels.
  • * Demonstrated multilineage differentiation potential and gene editing compatibility, with RNA-seq revealing enhanced pluripotency gene expression and metabolic activity.

Conclusions:

  • * The CD163-KO iPSC line is a validated, reliable in vitro model for PRRSV research.
  • * This cell line facilitates in-depth investigation of PRRSV entry mechanisms and host-pathogen interactions.
  • * The study provides a foundational tool for developing novel antiviral therapies and controlling porcine viral diseases.