Various fates of neuronal progenitor cells observed on several different chemical functional groups

doi:10.1007/s11706-011-0150-4

Front Mater Sci

2011, Vol. 5

Issue (4) : 358-366 https://doi.org/10.1007/s11706-011-0150-4

RESEARCH ARTICLE

Various fates of neuronal progenitor cells observed on several different chemical functional groups

Xi LIU¹, Ying WANG^1,2, Jin HE¹, Xiu-Mei WANG¹, Fu-Zhai CUI¹(

), Quan-Yuan XU²(

)

1. State Key Laboratory of New Ceramics and Fine Processing, Department of Materials Science and Engineering, Tsinghua University, Beijing 100084, China; 2. Beijing Institute for Neuroscience, Capital Medical University, Beijing Center of Neural Regeneration and Repair, Key Laboratory of Neurodegenerative Disease of Ministry of Education, Beijing 100069, China

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Abstract

Neuronal progenitor cells cultured on gold-coated glass surfaces modified by different chemical functional groups, including hydroxyl (-OH), carboxyl (-COOH), amino (-NH₂), bromo (-Br), mercapto (-SH), -Phenyl and methyl (-CH₃), were studied here to investigate the influence of surface chemistry on the cells’ adhesion, morphology, proliferation and functional gene expression. Focal adhesion staining indicated in the initial culture stage cells exhibited morphological changes in response to different chemical functional groups. Cells cultured on -NH₂ grafted surface displayed focal adhesion plaque and flattened morphology and had the largest contact area. However, their counter parts on -CH₃ grafted surface displayed no focal adhesion and rounded morphology and had the smallest contact area. After 6 days culture, the proliferation trend was as follows: -NH₂>-SH>-COOH>-Phenyl>-Br>-OH>-CH₃. To determine the neural functional properties of the cells affected by surface chemistry, the expression of glutamate decarboxylase (GAD67), nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) were characterized. An increase of GAD67 expression was observed on -NH₂, -COOH and -SH grafted surfaces, while no increase in NGF and BDNF expression was observed on any chemical surfaces. These results highlight the importance of surface chemistry in the fate determination of neuronal progenitor cells, and suggest that surface chemistry must be considered in the design of biomaterials for neural tissue engineering.

Keywords neuronal progenitor cell cell adhesion surface chemistry GABAergic neurotrophic factor

Corresponding Author(s): CUI Fu-Zhai,Email:cuifz@mail.tsinghua.edu.cn (F.Z.C.); XU Quan-Yuan,Email:xuqy@ccmu.edu.cn (Q.Y.X.)

Issue Date: 05 December 2011

Cite this article:

Xi LIU,Ying WANG,Jin HE, et al. Various fates of neuronal progenitor cells observed on several different chemical functional groups[J]. Front Mater Sci, 2011, 5(4): 358-366.

URL:

https://academic.hep.com.cn/foms/EN/10.1007/s11706-011-0150-4
https://academic.hep.com.cn/foms/EN/Y2011/V5/I4/358

Tab.1 Sequence of primers

Fig.1 Contact angle measurement. Results of mean±S.D. for five independent experiments ( = 5).

Fig.2 Confocal fluorescence microscopy images of focal adhesion and actin cytoskeleton in neuronal progenitor cells on chemical functional groups (incubation 4 h). Nuclear counterstaining revealed with DAPI (blue), focal contacts were revealed using anti-Vinculin monoclonal antibody (green), F-actin was detected using TRITC-conjugated phalloidin (red).

Fig.3 CCK-8 assay of neuronal progenitor cells cultured on different chemical functional groups after 2, 4, and 6 d. Symbol denotes significant differences: 1 symbol, <0.05; 2 symbols, <0.01. ★: compared to NH group on day 2; ◆: compared to NH group on day 4; ☆: compared to NH group on day 6. ( = 5)

Fig.4 Fluorescence microscopy of neuronal progenitor cells morphology on different chemical functional groups on day 1 and day 6. Fluorescent staining with Rhodamine phalloidin for F-actin (red) and SYTOXGreen for nuclei (green) showed the cell attachments and viabilities.

Fig.5 mRNA relative expression of GAD67, NGF and BDNF by quantitative RT-PCR after 6 d culture. Symbol denotes significant differences: 1 symbol, <0.05; 2 symbols, <0.01. ?: GAD67 expression compared to NH group; &: NGF expression compared to NH group; ◇: BDNF expression compared to NH group. ( = 3)

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