Establishing a 3D culture system for early organogenesis of monkey embryos ex vivo and single-cell transcriptome analysis of cultured embryos

Nianqin Sun; Honglian Shao; Youyue Zhang; Baiquan Ci; Hui Yao; Bing Bai; Tao Tan

doi:10.1016/j.xpro.2023.102835

Establishing a 3D culture system for early organogenesis of monkey embryos ex vivo and single-cell transcriptome analysis of cultured embryos

STAR Protoc. 2024 Mar 15;5(1):102835. doi: 10.1016/j.xpro.2023.102835. Epub 2024 Jan 14.

Authors

Nianqin Sun¹, Honglian Shao², Youyue Zhang², Baiquan Ci², Hui Yao², Bing Bai³, Tao Tan⁴

Affiliations

¹ State Key Laboratory of Primate Biomedical Research, Kunming University of Science and Technology, Kunming, Yunnan 650500, China; Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan 650500, China; Yunnan Key Laboratory of Primate Biomedical Research, Kunming, Yunnan 650500, China. Electronic address: [email protected].
² State Key Laboratory of Primate Biomedical Research, Kunming University of Science and Technology, Kunming, Yunnan 650500, China; Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan 650500, China; Yunnan Key Laboratory of Primate Biomedical Research, Kunming, Yunnan 650500, China.
³ State Key Laboratory of Primate Biomedical Research, Kunming University of Science and Technology, Kunming, Yunnan 650500, China; Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan 650500, China; Yunnan Key Laboratory of Primate Biomedical Research, Kunming, Yunnan 650500, China. Electronic address: [email protected].
⁴ State Key Laboratory of Primate Biomedical Research, Kunming University of Science and Technology, Kunming, Yunnan 650500, China; Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan 650500, China; Yunnan Key Laboratory of Primate Biomedical Research, Kunming, Yunnan 650500, China. Electronic address: [email protected].

Abstract

Creating in vitro culture platforms for monkey embryos is crucial for understanding the initial 4 weeks of early primate embryogenesis. Here, we present a protocol to culture cynomolgus monkey embryos in vitro for 25 days post-fertilization and to delineate the key developmental events of gastrulation and early organogenesis. We describe steps for culturing with a 3D system, immunofluorescence analysis, single-cell RNA sequencing, and bioinformatic analysis. For complete details on the use and execution of this protocol, please refer to Gong et al. (2023).¹.

Keywords: Bioinformatics; Cell Biology; Cell culture; Cell isolation; Developmental biology; Microscopy; Model Organisms; Molecular Biology; RNA-seq; Sequence analysis; Sequencing; Single Cell.

MeSH terms

Animals
Computational Biology
Embryonic Development / genetics
Macaca fascicularis
Organogenesis* / genetics
Single-Cell Gene Expression Analysis*