2. Academic Validation
  3. An efficient and user-friendly method for cytohistological analysis of organoids

An efficient and user-friendly method for cytohistological analysis of organoids

  • J Tissue Eng Regen Med. 2021 Nov;15(11):1012-1022. doi: 10.1002/term.3248.
Shi-Wei Zhang 1,  Wei Chen 2,  Xiao-Fang Lu 2,  Zhang Wen 2,  Lei Hu 2,  Yu-Hong Liu 3,  Zheng Yang 2,  Ling Xue 2,  Qiao Su 4,  Le-Ping Yan 1 5,  Joaquim M Oliveira 6 7,  Rui L Reis 6 7,  Yu-Long He 1,  Chang-Hua Zhang 1
Affiliations

Affiliations

  • 1 Guangdong Provincial Key Laboratory of Digestive Cancer Research, Digestive Diseases Center, The Seventh Affiliated Hospital of Sun Yat-Sen University, Shenzhen, China.
  • 2 Department of Pathology, The Seventh Affiliated Hospital of Sun Yat-Sen University, Shenzhen, China.
  • 3 General Surgery, The Seventh Affiliated Hospital of Sun Yat-Sen University, Shenzhen, China.
  • 4 Experimental Animal Center, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China.
  • 5 Scientific Research Center, The Seventh Affiliated Hospital of Sun Yat-Sen University, Shenzhen, China.
  • 6 3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Guimarães, Portugal.
  • 7 ICVS/3B's - PT Government Associate Laboratory, Guimarães, Portugal.
PMID: 34555270 DOI: 10.1002/term.3248
Abstract

Organoid culture is a recently developed in vitro three-dimensional (3D) Cell Culture technology. It has wide applications in tissue engineering studies. However, histological analysis of Organoid is quite complex and tedious for researchers. This study proposes a user-friendly, affordable and efficient method for making formalin-fixed paraffin embedded (FFPE) Organoid blocks and Optimal Cutting Temperature compound (OCT) embedded frozen Organoid blocks. This method implements a key pre-embedding step for preparing paraffin embedded Organoid blocks, which could concentrate Organoid together without damaging or loss of samples. This method could be used to process even a small number of organoids with high efficiency. In addition, with minor modifications, the method is readily applied for OCT embedded Organoid blocks. The slides generated were ready for H&E staining, immunohistochemistry staining and immunofluorescent staining. The method described in this study can be easily used for routine histological analysis of Organoid, and could be performed in general pathology labs and requires no dedicated equipment and reagent.

Keywords

FFPE; OCT; immunofluorescence; immunohistochemistry; organoids analysis.

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