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1、1About the role of laminin and type IV collagen in cultured nerve cells of the spinal cord neurite outgrowthAuthor: Han Yan, LU Kai, Yang Li, Zhang Hui Keywords laminin Abstract AIM: To detect the influence of laminin and type IV collagen on the axon growth of the cultured spinal neuron. METHODS: La
2、minin and collagen IV were used in the experimental group and laminin and collagen IV combined with associated antibody, collagen I and polylysine were used in the control groups. Spinal neuron cells of the same density were cultured in each group. Each group was stained by the immunochemical method
3、 of Neuron-specific Enolase (NSE) and the length of the neuron axons was measured by the computer image system. RESULTS: The spinal neuron cells and their axons were stained by NSE. The axons length of the laminin and collagen IV groups was longer than that of other groups but no significant differe
4、nce was observed. CONCLUSION: Laminin and collagen IV can stimulate the axon growth of spinal neuron cells. 2Keywords laminin, collagen, Neurons Abstract Objective: To observe laminin and type IV collagen role in neurite outgrowth of spinal cord nerve cells under in vitro culture conditions. Methods
5、: laminin and collagen type IV bag culture plates to combine corresponding antibody plates coated plates of type I collagen and polylysine control, the culture of spinal cord cells added to the plates at the same concentration. cultured cells in each group after 5 d neuron specific enolase enzyme (N
6、SE immunohistochemical staining using computer image analysis techniques groups neurite length and analyzed statistically. Results: Immunohistochemical staining is visible in each group have more nerve cells and their neurite outgrowth, statistical analysis results table BenQ laminin group and type
7、IV collagen group neurite growth length no significant difference in the laminin Group neurite growth length is longer than the other groups. collagen type IV group neuron axon growth length and type I the collagen group no significant differences between groups with poly-L-lysine significant differ
8、ence between the group of 3laminin and type IV collagen group neuron axon growth better than the corresponding antibody group Conclusion: laminin and IV The type of collagen can promote the growth of the spinal cord neuronal processes. Keywords laminin protein; collagen; neurons 0 Introduction Extra
9、cellular matrix components can promote the growth of the axons of nerve cells and myelin formation 1. Laminin and type IV collagen is an important component of the extracellular matrix, growth, development, and other aspects of the nerve cells plays an important The role of 2,3. observed in animal s
10、tudies of the two substances can promote the regeneration of peripheral nerve 4, is to learn more about the direct effect on neurite outgrowth of nerve cells from the morphology observed laminin and type IV collagen in cultured spinal cord nerve cells under the condition of neurite outgrowth. 1 Mate
11、rials and methods 1.1 Experimental groups were divided into six groups: laminin Group (LN 4group, anti-basement membrane adhesion the proteome (antiLN group of type IV collagen group (Col group, anti-collagen type IV group (antiCol group, I collagen type group (Col group, the group of poly-L-lysine
12、(poly group. Each group using the corresponding substrate package plates LN group: press 1 g cm-2, the amount of the base to laminin (GIBCO / BRL) is evenly applied to the culture plate. AntiLN group: packets are good LN group culture plates, anti-laminin monoclonal antibody (Beijing Zhongshan Biote
13、chnology Co., Ltd., working concentration 1:2000,37 overnight incubation. (3) of Col group: press 1 g cm-2 the amount of collagen (GIBCO / BRL evenly applied to the culture plate. AntiCol IV Group: Col group package is good training to join the anti-type IV collagen protein within the board monoclon
14、al antibody (Beijing Zhongshan Biotechnology Co., Ltd., work concentration 1:2000,37 overnight incubation. Col I Group: 1 g cm-2 the amount of type I collagen (Sigma evenly applied to the culture plate. Poly Group: a concentration of 20 g 5mL -1, poly-L-lysine (Sigma evenly applied to the culture pl
15、ate. Will be the above plates placed in the 37 C incubator for 24 h standby. 1.2 spinal cord cells grouped cultured under sterile conditions, cut the gestational age of 18 d Kunming the mouse spinal Reference Yang Dali 5 reported cell culture, under a dissecting microscope, carefully peel the spinal
16、 cord, stripping clean blood film, cut crushed tissue to 1 mm 1 mm x 1 mm, 125 g L-1 trypsin (Sigma) digestion at 37 C for 25 min with complete culture medium (DMEM containing fetal bovine serum, horse serum each 100 mL L- 1, Sigma) to terminate digestion for 10 min. N1 culture liquid (DF12 culture medium containing insulin 5 mg L-1, the transferrin protein 5 g L-1, the sodium selenate 5 g L-1, prog