Extraction and purification of neonatal versus adult rat Schwann cells / 中国组织工程研究

ABSTRACT

BACKGROUND: Schwann cells are the seed cells of neural repair, and it is a key to harvest a large number of Schwann cells with high purity and activity. OBJECTIVE: To compare the in vitro culture, purification, and morphology of Schwann cells between neonatal and adult rats, and investigate a simple and feasible culture method to harvest high-purity Schwann cells. METHODS: Totally 30 Sprague-Dawley rats, comprising 20 neonatal (1-3 days after birth, neonatal group) and 10 adult (weighing 150-200 g, adult group) rats, were included. Following double-enzyme digestion and two incubations, Schwann cells were isolated and purified by differential attachment. Cell morphology and attaching speed were determined through the use of inverted microscope. Cells were counted and cell purity was calculated. Cell proliferative ability was detected by MTT microcolorimetry. Curves of cell proliferation in each group were depicted to determine proliferative speed. Schwann cells were identified by S-100 immunochemistry.RESULTS AND CONCLUSION: Compared with fibroblasts, neonatal rat Schwann cells exhibited faster, while adult rat Schwann cells showed slower, attaching speed. Both neonatal and adult groups yielded over 96% cell purity. MTT microcolorimetry results revealed that Schwann cells proliferated actively in neonatal and adult groups. Cell proliferative curves show that neonatal rat Schwann cells proliferated faster than adult rat Schwann cells (P < 0.05). S-100 immunochemistry results showed positive results in both groups. All these findings suggest that double-enzyme digestion and two incubations followed by differential attachment is a satisfactory method to harvest considerable Schwann cells with high purity and activity. Neonatal rat Schwann cells show stronger proliferative, attaching capacities than adult rat Schwann cells.