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BACKGROUND:Neuroblastoma is a common solid tumor in children. Pediatric tumors are little affected by environmental factors, but closely related to child development. The suspension method is an effective and reliable method to harvest neoplastic stem cel s. OBJECTIVE:To culture the cel clones of human neuroblastoma cel line SK-N-SH and to assess the biological properties of the cel clones. METHODS:Using the suspension method with no serum media, tumor cel clones were obtained. Immunofluorescence method was used to identify whether tumor cel clones exhibit stem cel properties. SK-N-SH neuroblastoma was labeled by luciferase, and tumor cel clones and tumor cel s were seeded onto the back of nude mice to monitor cel proliferative properties in nude mice using in vivo imaging. RESULTS AND CONCLUSION:Using the suspension culture method, SK-N-SH neuroblastoma cel s could successful y develop into cloning bal s. Under serum-free culture, cloning bal s were immunofluorescently used to detect molecular markers that showed strong positive expression. Cloning bal s subcutaneously implanted into nude mice showed the strong ability of self-renewal and differentiation as stem cel s. The cel clones cultured by the suspension method strongly expressed Nestin, but weakly expressed glial fibril ary acidic protein, neuron-specific tubulin, possessing stem cel characteristics and strong proliferation and metastasis in nude mice.
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BACKGROUND:To in vitro isolate neural stem cel s with high purity and uniform biological properties and to establish a complete set of neural stem cel culture system is the basis for neural stem cel research. OBJECTIVE:To establish an isolation and culture system for neural stem cel s from newborn mouse hippocampus, olfactory bulb and cortex and to analyze the biological properties of cel s. METHODS:Neural stem cel s were isolated from the hippocampus, olfactory bulb and cortex tissue of newborn Kunming mice by mechanical separation and trypsin digestion. Serum-free culture technology, mechanical pipetting and trypsin digestion were used for subculture of neural stem cel s. 10%fetal bovine serum was used to induce differentiation of neural stem cel s. Neural stem cel s and their differentiated products were identified by immunofluorescent staining of Nestin, CD133,β-TubulinIII, glial fibril ary acidic protein. RESULTS AND CONCLUSION:The neural stem cel obtained from newborn mouse hippocampus, olfactory bulb and cortex had the capacity of self-renewal and differentiation which were positive for Nestin and CD133. After induction with fetal bovine serum, neural stem cel could differentiation toβ-tubulinIII or glial fibril ary acidic protein positive cel s that were neurons and astrocytes. This experiment has successful y established the neural stem cel isolation, culture, identification and induction system, providing experimental basis for subsequent studies of neural stem cel s.
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BACKGROUND:With the development of stem cel culture technology, stem cel transplantation has been a research hotspot in the biological cytology. OBJECTIVE:To review the current situation and progress of basic research on stem cel transplantation for human refractory diseases. METHODS:A computer-based search of CNKI, VIP, Wanfang and PubMed databases was performed for articles related to animal experiments of stem cel transplantation published from January 2008 to October 2014. The key words were“induced pluripotent stem cel s, neural stem cel s, top-up between stem cel s, transplantation”in Chinese and English in the title and abstract. Papers published in authoritative journals or recently were preferred. Total y 113 articles were searched initial y, and only 50 articles were included in result analysis. RESULTS AND CONCLUSION:With the progress of stem cel culture technology, basic research on the stem cel transplantation for human diseases have been promoted and carried out, and these researches have achieved phased outcomes that provide conditions for more extensive and in-depth fundamental research. It is believed that with the development of basic research on stem cel transplantation to solve a series of difficult problems, stem cel transplantation wil bring hope for treatment of human disease, especial y for treatment of refractory disease.
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10.3969/j.issn.2095-4344.2013.23.018