Scalable Production and Cryostorage of Organoids Using Core–Shell Decoupled Hydrogel Capsules

Abstract

Organoids, organ-mimicking multicellular structures derived from pluripotent stem cells or organ progenitors, have recently emerged as an important system for both studies of stem cell biology and development of potential therapeutics; however, a large-scale culture of organoids and cryopreservation for whole organoids, a prerequisite for their industrial and clinical applications, has remained a challenge. Current organoid culture systems relying on embedding the stem or progenitor cells in bulk extracellular matrix (ECM) hydrogels (e.g., Matrigel™) have limited surface area for mass transfer and are not suitable for large-scale productions. Here, we demonstrate a capsule-based, scalable organoid production and cryopreservation platform. The capsules have a core-shell structure where the core consists of Matrigel™ that supports the growth of organoids, and the alginate shell form robust spherical capsules, enabling suspension culture in stirred bioreactors. Compared with conventional, bulk ECM hydrogels, the capsules, which could be produced continuously by a two-fluidic electrostatic co-spraying method, provided better mass transfer through both diffusion and convection. The core-shell structure of the capsules also leads to better cell recovery after cryopreservation of organoids probably through prevention of intracellular ice formation.

Vitamin C and B3 as new biomaterials to alter intestinal stem cells

“…Thus, the organoids have identical cell pathways and similar cell types when compared to the intestine epithelium itself. These advantages have been used to study various topics, such as modeling infectious diseases (Liu, Huang, Ning, et al, ), applications to different drug delivery systems (Cheng et al, ; Peng, Wang, Xu, Yu, & Wang, ; Tu et al, ; Wang et al, ; Wu, Deng, Jiang, et al, ; Zhao, Wang, Zhang, et al, ), how they serve as a new drug loading material (Davoudi et al, ), various studies on nanomaterial's toxicity (Qi et al, ), modeling different virus infections (Garcez, Loiola, Da Costa, et al, ; Yin et al, ), studying microbiota interactions (Lukovac et al, ), and many more (Chiu, Song, Flanders, et al, ; Lu et al, ; Song et al, ; Zhang, An, Pardo, et al, ).…”

Advanced Biotechnology for Cell Cryopreservation

“…3d). The core-shell structure of the capsules provided better cell recovery after cryopreservation of organoids, probably through prevention of intracellular ice formation [73][74][75]. Electrohydrodynamic atomization (EHDA) is an attractive approach that immobilizes living cells into biomaterials permitting localized and minimally invasive delivery.…”

Exploiting CRISPR Cas9 in Three-Dimensional Stem Cell Cultures to Model Disease

“…Polysaccharides like alginate have also been shown to support growth of hiPSC-derived and patient-specific organoids, rendering similar phenotypic traits comparable to Matrigel-grown organoids ( Lu et al, 2017 ; Wilkinson et al, 2017 ; Broguiere et al, 2018 ; Capeling et al, 2019 ). Osteogenesis and angiogenesis could be achieved in an artificial sphere-shaped organoid using mesenchymal stem cells (MSCs) and human umbilical vain endothelial cells (HUVECs) ( Zhao et al, 2018 ).…”