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Biology subjects

Hillenius, S.

Publications and source records attributed to Hillenius, S..

2 recordsLinked to original sources

Cytogenetic resource enables mechanistic resolution of changing trends in human pluripotent stem cell aberrations linked to feeder-free culture

Since the first derivation of human pluripotent stem cells (hPSCs), the number of culture conditions has steadily increased, making hPSC culture more facile. Nonetheless, there remains the persistent issue of culture-acquired genetic changes, hampering the reproducibility of hPSC research and jeopardising their clinical use. Here, we utilised comprehensive karyotyping datasets from over 20,000 hPSC cultures sampled under different conditions to ascertain association of genetic changes with specific culture regimens. We found condition-dependent patterns of aberrations, with higher prevalence of chromosome 1q gains in recent years, associated with increased use of contemporary, feeder-free cultures. Mechanistically, we show the context-dependent selection of 1q variants is mainly driven by MDM4, a gene amplified in many cancers, located on chromosome 1q. To facilitate reproducibility of hPSC research and their safe clinical utility, we provide a unique hPSC karyotype resource for informing the risk assessment of genetic aberrations and developing strategies to suppress their occurrence.

cell biology↗

Efficient and scalable generation of primordial germ cells in 2D culture using basement membrane extract overlay

Current human primordial germ cell like cells (hPGCLCs) differentiation methods from human pluripotent stem cells (hPSCs) are inefficient, and it is challenging to generate sufficient hPGCLCs to optimize in vitro gametogenesis. We present a new differentiation method that uses diluted basement membrane extract (BME) and low BMP4 concentration to efficiently induce hPGCLC differentiation in scalable 2D cell culture. We show that BME overlay potentiated BMP/SMAD signaling, induced lumenogenesis and increased expression of key hPGCLC-progenitor markers such as TFAP2A and EOMES. These findings highlight the importance of (factors in) BME during hPGCLC differentiation, and demonstrate the potential of the BME-overlay method to interrogate the formation of PGCs and amnion in humans as well to investigate the next steps to achieve in vitro gametogenesis. HIGHLIGHTSO_LIhPGCLCs can be generated efficiently in 2D from hPSCs with treatment of BMP4 and BME overlay C_LIO_LIBME overlay method is highly scalable, cost-effective and simple to perform C_LIO_LIhPGCLCs differentiate together with amniotic ectoderm- and mesoderm-like cells from a TFAP2A+/CDX2+/EOMES+/GATA3+ common progenitor population C_LIO_LIBME overlay enables robust hPGCLC formation by potentiating BMP/SMAD signaling in the common progenitor population C_LI

developmental biology↗