Culture of Neural Stem Cells in Calcium Alginate Beads

Abstract

Neural stem cells (NSCs) with the capacity of extensive self-renewal and multilineage differentiation have attracted more and more attention in research as NSCs will play an important role in the nerve disease treatment and nerve injury repair. The shortage of NSCs, both their sources and their numbers, however, is the biggest challenge for their clinic application, and hence, 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 exploration of formation and dissociation parameters for Ca-Alg-Bs. Additionally, the recovered and the subsequent induced cells were identified by immunofluorescence staining of Nestin, beta-tubulin, and GFAP. The results show that the 2-mm diameter Ca-Alg-Bs, prepared with 1.5% sodium alginate solution and 3.5% CaCl2 solution and with gelling for 10 min, is suitable for the NSCs culture. The seeding density of 0.8 x 10(5) cells x mL-1 for the encapsulation of NSCs resulted in the most expansion, and the NSCs almost doubled during the experiment. The average cell recovery rate is over 88.5%, with the Ca-Alg-Bs dissolving in 55 mM sodium citrate solution for 10 min. The recovered cells cultured in the Ca-Alg-Bs still expressed Nestin and had the capacity of multilineage differentiation into neurons and glial cells and, thus, remained to be NSCs. These results demonstrate that NSC expansion within Ca-Alg-Bs is feasible and provides further possibilities for NSC expansion in bioreactors of the scale of clinical relevance.

Hydrogels from Biopolymer Hybrid for Biomedical, Food, and Functional Food Applications

“…Although alginate lacks a specific cell-recognition site, the introduction of specific cell adhesive motifs into the structure of alginate-based biohydrogels imparts the ability to react specifically with mammalian cells or function as an artificial extracellular matrix to promote cell adhesion, proliferation and guide cell differentiation. This further broadens the applications of alginate biohydrogels in regenerative medicine into areas such as wound healing, bone and cartilage regeneration, construction of blood vessels, and many others [10,[20][21][22][23][24][25][26]. The injection of in situ-forming, bioabsorbable alginate hydrogels has been shown to be an effective acellular strategy that prevents adverse cardiac remodeling and dysfunction in new and old myocardial infarction in rats [27].…”

Alginate Composition Effects on a Neural Stem Cell–Seeded Scaffold

“…Recent studies have investigated the use of alginate as a neural stem cell (NSC) scaffold. [5][6][7] However, the effect of alginate composition on NSC function has not been investigated.…”

Technological progresses in monoclonal antibody production systems

“…63 The most wellknown system for culture of adherent cells is microcarrier culture, 64,65 which can be performed in different bioreactors, with STR being the most common. 66 Other immobilization systems include small beads of different materials 67 (agarose, 68 gelatin and alginate 67,69 ), gel particles (collagen 70 ), membranes, [71][72][73] and fibers. [74][75][76][77] All these systems are ideal for perfusion culture, 11 which increases cell concentration in the reactor (10 7 -10 8 cells/mL) and extends the productive lifetime of cells, 60,74 resulting in increased levels of mAb productivity when compared with suspension culture.…”