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	Isolation, culture, and purification of olfactory mucosa-derived olfactory ensheathing cells using modified differential attachment with low concentration serum
	Yang, Huaqing ; Li, Qiang ; Wang, Kunzheng ; Wang, Bin ; Qiang, Hui ; Wang, Wei ; Yao, Jianxiang
	2010-05-11 ; 2010-05-11
关键词	cell culture olfactory ensheathing cells olfactory mucosa purification SCHWANN-CELLS ADULT-RAT NEUROTROPHIN-3 NEURONS TRANSPLANTATION RECOVERY Cell Biology Neurosciences
中文摘要	BACKGROUND: Studies have demonstrated that olfactory mucosa call promote the regeneration and jured neurons. To date, olfactory ensheathing cells (OECs) utilized formation of axonal medullary sheath of in. in basic aid clinical research arise primarily from the olfactory bulb mucosa. However, little is known regarding Culture, purification, and biological properties of OECs. OBJECTIVE: To isolate and culture OECs utilized modified, differential attachment in combination with neurotrophic factor 3 (NT3) and low concentration Serum to explore an optimal in vitro Culture method for OECs. DESIGN, TIME AND SETTING: Single-sample observation was performed at the Medical Experimental Center of Stomatology College, Xi'an Jiaotong University between March 2006 and December 2007. MATERIALS: Twelve samples from aborted embryo,,, 44) months, were used to isolate OECs; rabbit-anti-human p75(NTR) and glial fibrillary acidic protein (GFAP) antibody were provided by Sigma, USA. METHODS: The differential time was six hours. This was repeated twice, based on Nash's differential attachment. Attached OECs were cultured in DMEM-F-12 culture medium containing 10% fetal bovine serum (FBS) or 2.5% FBS and NT3. MAIN OUTCOME MEASURES: OEC morphology was observed, and p75(NTR) and GFAP immunocytochemistry was used for identification and purity detection. RESULTS: Some cells attached after three days in culture. Several cells possessed short neurites with good refractivity. Some shuttle-shaped fibroblasts could be seen. Oil day six, more cells attached, exhibiting a three-dimensional appearance. Many cells appeared dipolar or tripolar, with slender neurites, and fibroblasts were clustered. On day nine, the number of dipolar or tripolar cell bodies with slender neurites was increased, and fibroblasts were clustered. Oil day 15, fibroblasts occupied the majority of the bottom of the culture bottle, with several OECs Surviving at the upper layer. OECs were positive for P75(NTR) and GFAP expression, as identified by all immunocytologically stained brown cell body and neurites. However, fibroblasts were P75(NTR) and GFAP-negative. On day 9, OEC purity reached 81%, and the number of proliferating fibroblasts significantly increased. By the end of day 12, OEC purity was reduced to 56%. CONCLUSION: Modified differential attachment, in combination with low concentration serum and NT3, removes fibroblasts and reduces OEC loss. This is an appropriate method for the isolation and culture of human fetal olfactory mucosa-derived OECs.
语种	英语 ; 英语
出版者	SHENYANG EDITORIAL DEPT NEURAL REGENERATION RES ; SHENYANG ; PO BOX 1234, SHENYANG, LIAONING 110004, PEOPLES R CHINA
内容类型	期刊论文
源URL	[http://hdl.handle.net/123456789/27773]
专题	清华大学
推荐引用方式 GB/T 7714	Yang, Huaqing,Li, Qiang,Wang, Kunzheng,et al. Isolation, culture, and purification of olfactory mucosa-derived olfactory ensheathing cells using modified differential attachment with low concentration serum[J],2010, 2010.
APA	Yang, Huaqing.,Li, Qiang.,Wang, Kunzheng.,Wang, Bin.,Qiang, Hui.,...&Yao, Jianxiang.(2010).Isolation, culture, and purification of olfactory mucosa-derived olfactory ensheathing cells using modified differential attachment with low concentration serum..
MLA	Yang, Huaqing,et al."Isolation, culture, and purification of olfactory mucosa-derived olfactory ensheathing cells using modified differential attachment with low concentration serum".(2010).

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