2. Academic Validation
  3. Derivation of Haploid Neural Stem Cell Lines by Selection for a Pax6-GFP Reporter

Derivation of Haploid Neural Stem Cell Lines by Selection for a Pax6-GFP Reporter

  • Stem Cells Dev. 2018 Apr 1;27(7):479-487. doi: 10.1089/scd.2017.0193.
Qian Gao 1 2,  Wenhao Zhang 1,  Lifang Ma 1,  Xu Li 1,  Haisong Wang 1,  Yanni Li 1,  Remo Freimann 3,  Yang Yu 2,  Ling Shuai 1,  Anton Wutz 3
Affiliations

Affiliations

  • 1 1 State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Nankai University , Tianjin, China .
  • 2 2 Reproductive Medical Center, Department of Gynecology and Obstetrics, Peking University Third Hospital , Beijing, China .
  • 3 3 Institute of Molecular Health Sciences, Swiss Federal Institute of Technology Zurich , Zurich, Switzerland .
PMID: 29471728 DOI: 10.1089/scd.2017.0193
Abstract

Haploid cells facilitate genetic screening of recessive mutations for a single set of chromosomes. Haploid embryonic stem cells (haESCs) have been achieved in several species and widely utilized in genetic screens. The fact that haESCs undergo substantial diploidization during differentiation has limited the screening to other haploid cell types. In this study, we report a method to establish haploid neural stem cells (haNSCs) by selection for a Pax6 reporter. We inserted a green fluorescence protein (GFP) marker gene by homologous recombination into the Pax6 locus of an haESC line. GFP-positive haploid cells could be sorted and further cultured in the NSC medium for more than 30 passages. The established haNSCs expressed neural lineage markers and could differentiate into neurons, oligodendroglia, and astrocytes. Our study shows the feasibility of deriving haploid proliferative somatic cell lines using a genetically encoded reporter that suggest a system for genetic screening of neural and retinal development.

Keywords

CRISPR; Pax6-GFP; haploid; neural stem cells; nontoxic sorting.

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