Amsterdam University Medical Center

2 ARTICLES PUBLISHED IN JoVE

Bioengineering

Generation, High-Throughput Screening, and Biobanking of Human-Induced Pluripotent Stem Cell-Derived Cardiac Spheroids

Renee G. C. Maas¹, Tess Beekink^*1, Nino Chirico^*1, Christian J. B. Snijders Blok¹, Inge Dokter¹, Vasco Sampaio-Pinto¹, Alain van Mil¹, Pieter A. Doevendans¹, Jan W. Buikema², Joost P. G. Sluijter^*1, Francesca Stillitano^*1

¹Utrecht Regenerative Medicine Center, Circulatory Health Laboratory, University Utrecht, Department of Cardiology, University Medical Center Utrecht, ²Amsterdam Cardiovascular Sciences, Department of Physiology, Amsterdam University Medical Center

Presented here is a set of protocols for the generation and cryopreservation of cardiac spheroids (CSs) from human-induced pluripotent stem cell-derived cardiomyocytes cultured in a high-throughput, multidimensional format. This three-dimensional model functions as a robust platform for disease modeling, high-throughput screenings, and maintains its functionality after cryopreservation.

Developmental Biology

Human Ovarian Surface Epithelium Organoids as a Platform to Study Tissue Regeneration

Julieta S. Del Valle^1,2, Azra Husetic^*1, Dina Diek^*1, Laurens F. Rutgers^*1, Joyce D. Asseler^3,4,5, Jeroen Metzemaekers⁶, Norah M. van Mello^3,4,5, Susana M. Chuva de Sousa Lopes^1,2,7

¹Department of Anatomy and Embryology, Leiden University Medical Center, ²The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center, ³Department of Obstetrics and Gynaecology, Amsterdam University Medical Center, ⁴Centre of Expertise on Gender Dysphoria, Amsterdam UMC, ⁵Amsterdam Reproduction and Development Research Institute, ⁶Department of Gynaecology, Leiden University Medical Center, ⁷Ghent-Fertility and Stem Cell Team (G-FAST), Department of Reproductive Medicine, Ghent University Hospital

This protocol describes establishing three-dimensional (3D) tissue organoids from primary human ovarian surface epithelium (hOSE) cells. The protocol includes isolation of hOSE from freshly collected ovaries, cellular expansion of the hOSE, cryopreservation-thawing procedures, and organoid derivation. Immunofluorescence, quantitative analysis, and showcasing utility as a screening platform are included.