The authors stated that the absence of data on the embryonic development of the colon is partly to blame as an obstacle against the ability to properly identify the right molecular and genetic programming necessary to get the human pluripotent stem cells into the colonic organoids. The researchers overcame this obstacle by taking the developing hindgut tissues out of animal models and using it to create a series of genetic and molecular screens. The entire large intestine (cecum, rectum, and colon) is given rise from the hindgut, which is a part of the developing gut.
The team also identified colon hindgut molecular markers by mining public databases, such as GNC Pro, TiGER, and the Human Protein Atlas.
The scientists used frogs and mice to identify the SATB2 (special AT-rich sequence-binding protein 2) as a defining molecular marker for hindgut which allowed for the development of the human colon generating model.