Culture of Neural Stem Cells in Calcium Alginate Beads

详细信息    查看全文

• 作者：Xiangqin Li ; Tianqing Liu ; Kedong Song ; Lisong Yao ; Dan Ge ; Chunyu Bao ; Xuehu Ma ; Zhanfeng Cui
• 刊名：Biotechnology Progress
• 出版年：2006
• 出版时间：December 2006
• 年：2006
• 卷：22
• 期：6
• 页码：1683 - 1689
• 全文大小：485K
• 年卷期：v.22,no.6(December 2006)
• ISSN：1520-6033

文摘

Neural stem cells (NSCs) with the capacity of extensive self-renewal and multilineagedifferentiation have attracted more and more attention in research as NSCs will play an importantrole in the nerve disease treatment and nerve injury repair. The shortage of NSCs, both theirsources and their numbers, however, is the biggest challenge for their clinic application, andhence, in vitro culture and expansion of NSCs is vitally important to realize their potentials.In this work, mouse-derived NSCs were cultured in three-dimensional calcium alginate beads(Ca-Alg-Bs). Gelling conditions, cell density, and cell harvest were determined by the explorationof formation and dissociation parameters for Ca-Alg-Bs. Additionally, the recovered and thesubsequent induced cells were identified by immunofluorescence staining of Nestin, -tubulin,and GFAP. The results show that the 2-mm diameter Ca-Alg-Bs, prepared with 1.5% sodiumalginate solution and 3.5% CaCl₂ solution and with gelling for 10 min, is suitable for the NSCsculture. The seeding density of 0.8 × 10⁵ cells·mL^-1 for the encapsulation of NSCs resulted inthe most expansion, and the NSCs almost doubled during the experiment. The average cellrecovery rate is over 88.5%, with the Ca-Alg-Bs dissolving in 55 mM sodium citrate solutionfor 10 min. The recovered cells cultured in the Ca-Alg-Bs still expressed Nestin and had thecapacity of multilineage differentiation into neurons and glial cells and, thus, remained to beNSCs. These results demonstrate that NSC expansion within Ca-Alg-Bs is feasible and providesfurther possibilities for NSC expansion in bioreactors of the scale of clinical relevance.