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Kidney Week

Abstract: TH-OR034

Single Bioreactor Culture System for Mass Production of Kidney Organoids

Session Information

  • Bioengineering
    November 07, 2019 | Location: 146 A/B, Walter E. Washington Convention Center
    Abstract Time: 05:06 PM - 05:18 PM

Category: Bioengineering

  • 300 Bioengineering

Authors

  • Matsumoto, Takuya, harvard medical school, Boston, Massachusetts, United States
  • Morizane, Ryuji, Brigham and Women's Hospital, Boston, Massachusetts, United States
Background

Pluripotent stem cell (PSC)-derived kidney organoids have great potential to recapitulate multicellular relationships and microenvironments of native kidneys, which can be used for drug screening, toxicology assays, disease modeling, and regenerative therapy. Mass production of kidney organoids by a robust, reproducible, and low labor-load approach is a prerequisite for the transition from laboratory to industrial applications. However, the process hasn’t been established yet. Here, we employ stirred bioreactor culture system to manufacture kidney organoids and demonstrate a method of mass production of organoids with a well-established quality control.

Methods

We used Biott stirred bioreactor system. Bioreactors were inoculated with single cell suspension of human PSCs at optimal cell density in StemFit® Basic02 in a final 5-ml culture volume. Then, kidney organoids were differentiated by 6-step growth factor treatment which were modified from our previously reported protocols. Induction of nephron progenitor cells (NPCs) and kidney organoids were validated using a combination of immunostaining and qPCR.

Results

500-1000 organoids were generated in a 5-ml bioreactor. Kidney organoids cultured in this system contained proximal tubular cells (LTL+), podocytes (NEPHRIN+), distal tubular cells (LTL-ECADHERIN+), interstitial cells (MEIS1/2+/ PDGFRβ+), and endothelial cells (CD31+). The sphere size at the beginning of differentiation was identified as a dominant factor which affected induction efficiency of NPCs and organoids. Optimization of CHIR concentration was also important for efficient NPC differentiation. Of note, higher expression of SIX2 at the NPC differentiation stage was positively correlated with more nephron structures in organoids which were viable for a longer time (>35 days) without apoptosis.

Conclusion

We established stirred bioreactor culture system for manufacturing kidney organoids whose yield was 10 times more efficient than the conventional culture system (96 organoids in 96 well culture plates). Our results also indicated SIX2 expression in NPCs is a predictive marker for production of high-quality kidney organoids. This process could be readily scaled up to support development of cell-products for clinical use at industry levels in the future.

Funding

  • NIDDK Support