Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Light Sheet Microscopy and 3D Analysis of Human iPSC-derived Embryoid Bodies

Mohammed Yusuf
* ,
Jahan Salma
* ,
Safana Farooq
ORCID logo ,
Alessia Candeo
,
Archana Bhartiya
,
Atiqa Sajid
,
Marta Szynkiewicz
,
Ian Robinson
ORCID logo ,
Stan Botchway
,
El-Nasir Lalani
Version 1 : Received: 28 December 2022 / Approved: 13 January 2023 / Online: 13 January 2023 (07:10:46 CET)

How to cite: Yusuf, M.; Salma, J.; Farooq, S.; Candeo, A.; Bhartiya, A.; Sajid, A.; Szynkiewicz, M.; Robinson, I.; Botchway, S.; Lalani, E. Light Sheet Microscopy and 3D Analysis of Human iPSC-derived Embryoid Bodies. Preprints 2023, 2023010238. https://doi.org/10.20944/preprints202301.0238.v1 Yusuf, M.; Salma, J.; Farooq, S.; Candeo, A.; Bhartiya, A.; Sajid, A.; Szynkiewicz, M.; Robinson, I.; Botchway, S.; Lalani, E. Light Sheet Microscopy and 3D Analysis of Human iPSC-derived Embryoid Bodies. Preprints 2023, 2023010238. https://doi.org/10.20944/preprints202301.0238.v1

Abstract

Embryoid bodies (EBs) are multicellular three-dimensional (3D) aggregates generated from induced pluripotent stem cells (iPSCs) in suspension and serve as useful biological sources for many downstream applications. Imaging of live EBs has been hampered mainly due to the inherent limitations of the imaging techniques applied to date. This study aimed to image human iPSC (hiPSC) derived EBs to obtain their 3D volume, determining size, morphology, and cell viability from day 7 to 14 using Light Sheet Fluorescence Microscopy (LSFM). Furthermore, chromosomal stability was assessed using Multicolor fluorescence in situ hybridization (M-FISH) from day 8 to 14. EB volume increased from day 7 to 13 which, decreased at day 14. From day 7 to 11, the EBs mainly appeared spherical and morphed into an ellipsoidal shape by day 13. All EBs showed varied external morphologies and larger cavities at day 14. The EB karyotype was diploid 46XY at day 8 and exhibited a low level of aneuploidy from day 10 to 14. This study shows that an increase in cell death affects the morphology and chromosomal stability in EBs derived from hiPSC. We demonstrate that the combination of LSFM and M-FISH helps characterize EBs that will assist future stem cell therapies.

Keywords

induced pluripotent stem cells; embryoid bodies; light-sheet microscopy; multicolor fluorescence in situ hybridization; chromosomes

Subject

Biology and Life Sciences, Cell and Developmental Biology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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