Isolation, Characterization and Transplant of intestinal-Derived Neural Stem Cells as a Treatment for Intestinal Neuropathies
Research of Jaime Belkind-Gerson, MD
Jaime Belkind-Gerson, MD
Staff Physician, Pediatric Gastroenterology & Nutrition, MassGeneral Hospital for Children; Instructor in Pediatrics, Harvard Medical School
The enteric nervous system (ENS) regulates important intestinal functions, including motility, sensation, absorption, secretion, blood flow, and immune function. When injured or lacking, intestinal function is severely compromised. We were one of the first groups to describe that the intestine contains neural stem cells (NSCs) capable of generating new neurons (neurogenesis) throughout adult life, opening up the potential for using autologous therapies to treat ENS disorders. Several others have corroborated the presence of intestinal NSCWe believe that successful isolation, expansion, and transplantation of NSCs will hasignificant impact on treatment of intestinal neuropathies and also help to clarify the mechanisms governing ENS regeneration and plasticity.
Intestinal NSCs were first identified as cells in the intestinal muscularis. We have isolated NSCs from cultures derived from intestinal muscularis, but also from the mucosa. The presence of a mucosal cell capable of giving rise to NSCs is further supported by Metzger at al (Gastroenterology 2009), who generated NSCs from mucosal endoscopic biopsies from children. This offers a minimally invasive and autologous source of NSCs for cell-based therapies in enteric neuropathies, including Hirschsprung’s disease. However, the identity of the intestinal particularly the mucosal
NSCs is unknown. Furthermore, mucosal biopsies are very small and contain few cells, making NSC isolation difficult. We are working to isolate and characterize mucosa-derived NSCs using immunosorting and other cell-enrichment techniques with the broad objectives of developing these cells as a source of neurons for the treatment of neurointestinal diseases. So far, in our attempts at characterization, we have grown mucosal-derived NSCs as both: a monolayer culture and in floating colonies called neurospheres (Figures 1 and 4). The NSCs, starting at day 4, differentiate into neurons and glia (Figure 2). The neurons resulting from this differentiation express several neurotransmitters normally found in the mucosa (Figure 3). We are currently grafting these NSCs into embryonic chick and quail hindguts and human aganglionic gut, to asses for survival, differentiation to neurons and glia and function.
Our overall goal is to continue to characterize both mucosal and muscularis-derived NSCs and to study their role in ENS regeneration in health and response to disease, as well as using the NSCs as possible cell-therapy for various forms of intestinal neuropathy including Hirschsprung’s disease.