In vivo evaluation of scaffolds compatible for colonoid engraftments onto injured mouse colon epithelium

Abstract

Colon organoids (colonoids) are known to be similar to colon tissue in structure and function, which makes them useful in the treatment of intestinal de‐epithelialized disease. Matrigel, which is used as a transplantation scaffold for colonoids, cannot be used in clinical applications because of its undefined composition and tumorigenicity. This study identifies clinically available scaffolds that are effective for colonoid transplantation in damaged intestinal mucosa. The colon crypt was isolated and cultured from C57BL/6‐Tg[CAG enhanced green fluorescent protein (EGFP)1310sb/LeySopJ mice into EGFP + colonoids and subsequently transplanted into the EDTA colitis mouse model using gelatin, collagen, or fibrin glue scaffolds. To identify scaffolds suitable for colonoid engraftment in injured colon mucosa, the success rates of transplantation and secondary EGFP colonoid formation were measured, and the scaffolds' mediated toxicity in vitro and in vivo was observed in recipient mice. When colonoids were transplanted with gelatin, collagen, and fibrin glue into the EDTA colitis mouse model, all groups were found to be successfully engrafted. Fibrin glue, especially, showed significant increase in the engrafted area compared with Matrigel after 4 wk. The scaffolds used in the study did not induce colonic toxicity after transplantation into the recipients' colons and were thus deemed safe when locally administrated. This study suggests new methods for and provides evidence of the safety and utility of the clinical application of colonoid‐based therapeutics. Furthermore, the methods introduced in this study will be helpful in developing cell treatment using the esophagus or a stomach organoid for various digestive‐system diseases.—Jee, J., Jeong, S. Y., Kim, H. K., Choi, S. Y., Jeong, S., Lee, J., Ko, J. S., Kim, M. S., Kwon, M.‐S., Yoo, J. In vivo evaluation of scaffolds compatible for colonoid engraftments onto injured mouse colon epithelium. FASEB J. 33, 10116–10125 (2019). http://www.fasebj.org

Biomaterials and biosensors in intestinal organoid culture, a progress review

“…Moreover, the stiffness of the fibrin‐based hydrogel can affect intestinal organoid differentiation. Usually, scaffold stiffness is regulated by altering polymer density (Jee et al, ). The fibrin at 3–4.5 mg/ml has a storage modulus of 80–140 Pa, which is the optimal mechanical strength for intestinal organoid culture (Broguiere et al, ).…”

Attenuation of Chronic Inflammation in Intestinal Organoids with Graphene Oxide-Mediated Tumor Necrosis Factor-α_Small Interfering RNA Delivery

“…50 Our organoid culture system aimed to replicate this intricate cellular structure and communication in an in vitro setting. To assess organotypic morphology, we conducted staining for key markers such as Lgr5, lysozyme, Muc2, ChgA, villin, and Ecadherin 51 (as illustrated in Figure 1B−C). Both enteroids and colonoids exhibited enterocytes expressing E-cadherin at the basolateral membrane and villin at the apical surface, akin to in vivo conditions.…”

Optimized protocol for the generation of an orthotopic colon cancer mouse model and metastasis

“…Alternative models include injecting colon cancer cells subcutaneously or in the cecum; however, that does not recapitulate the natural microenvironment for colon cancer cells. While several publications have recently described the transplantation of normal organoids into injured colonic mucosa, 1 , 2 here we describe a protocol that enables the generation of primary colonic tumors that can metastasize into the liver. This rapid and reproducible protocol also significantly reduces mouse mortality due to the procedure.…”