A fast and efficient method for isolating and culturing enteric neural precursor cells from adult mouse colon.

BACKGROUND:The enteric neural precursor cells (ENPCs) are important for researching the pathogenesis of enteric nervous system (ENS)-related diseases, especially in adulthood. Because primary ENPCs are difficult to isolate and survive, easy to contaminate and low-yielding, a rapid and effective method to isolate and cultivate ENPCs from adult mice is necessary. NEW METHODS:The longitudinal muscle myenteric plexus (LMMP) was isolated from the adult mouse colon. The papain and collagenase Ⅱ were chosen to increase the yield of ENPCs. The growth and proliferation of ENPCs could be promoted by using polylysine precoated culture plates and reasonable cell seeding density. The ENPCs were identified by Nestin and the proliferative properties were verified by EDU. The transgenic Nestin-cre:tdTomato mice were used to observe the proliferation of ENPCs more intuitively in vitro. RESULTS:Our method to isolate the ENPCs from adult mouse colon was effective and high-yielding. The ENPCs were identified as Nestin positive. The Nestin-positive ENPCs could proliferate rapidly and had a tendency to differentiate into neurons and glial cells in vitro without any inducing factors. COMPARISON WITH EXISTING METHODS:Previous studies about the ENPCs focused on experiment in vivo or were limited to the embryonic mice, and had limitations of low yield and long experiment time. The ENPCs from adult mice were isolated quickly by our method, and had a high yield and purity. CONCLUSION:We described a rapid, efficient, step by step method for isolation and culture of ENPCs from the adult mouse colon, which was simple and obtained high yield of ENPCs.