Generation of renal organoids from mouse embryonic stem cells

Thesis event information

Date and time of the thesis defence

Place of the thesis defence

Auditorium F202, Aapistie 5 B, Oulu, Finland

Topic of the dissertation

Generation of renal organoids from mouse embryonic stem cells

Doctoral candidate

Master of Cell Biology Zenglai Tan

Faculty and unit

University of Oulu Graduate School, Faculty of Biochemistry and Molecular Medicine, Developmental Biology

Subject of study

Biochemistry and molecular medicine

Opponent

Professor Nuria Montserrat, Institute for Bioengineering of Catalonia

Custos

Professor Seppo Vainio, Faculty of Biochemistry and Molecular Medicine

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Generation of renal organoids from mouse embryonic stem cells

The application of pluripotent stem cell (PSC) and organoid technologies provides new ways to study kidney disease and kidney development and opens up new possibilities for developing therapy strategies. This study presents new methods with which to generate renal cells from mouse embryonic stem cells (mESCs).

We first differentiated mESCs into nephron progenitor cells (NPCs). Then we aggregated the mESC-derived NPCs with mouse embryonic ureteric buds (UB) and transferred them to a three-dimensional (3D) organ culture system. Upon UB induction, the mESC-derived NPCs were able to undergo nephrogenesis and form appropriately segmented nephron structures. Wnt4 plays an essential role during nephrogenesis and Tcf21 is involved in the induction of nephrogenesis, we knocked out the Wnt4 and Tcf21 functions in the mESCs with clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated systems 9 (Cas9) and differentiated the mutant nephron progenitor cells to generate renal organoids. The Wnt4 CRISPR-mutant organoids failed to undergo nephrogenesis and lacked nephron structures. The Tcf21-deficient organoids showed cyst-formation in the organoids. We then optimized the protocol for direct differentiation of mESCs into UB progenitors. We generated kidney organoids by aggregating the UB progenitors with mouse embryonic metanephric mesenchyme (MM). In the chimeric kidney organoids, the embryonic MM developed into nephrons containing all segments. The mESC-derived UB progenitors formed a collecting duct system interconnected with the nephron tubules. The organoids generated from the mESC-derived UB progenitor cells presented toxicity to nephrotoxic drugs. This indicates that these organoids are good models to study renal development and toxicity.

In summary, we established new differentiation protocols to derive nephron and collecting duct progenitors, and used the kidney organoid platform to model kidney development, disease, and injury.
Last updated: 12.9.2019