The enhancement of neural stem cell survival and growth by coculturing with expanded sertoli cells in vitro

Abstract

Sertoli cells (SCs) have been described as the "nurse cells" of testis to provide essential growth factors and to create a proper environment for the development of other cells (e.g., germinal and neural stem cell). However, the physiological functions of the SCs obtained from different culture conditions are different in a coculturing system, and thus the optimal SC culturing condition should be investigated in vitro. In this paper, primary Sertoli cells were isolated from a 12-day-old mouse and expanded in two different culture conditions: a two dimensional (2D) plastic tissue disc and a three dimensional (3D) microcarrier culture system. They were then cocultured with neural stem cells (NSCs) isolated from 14-day-old mouse embryos. The metabolic activities of SCs(2D) (SCs in 2D) and SCs(3D) (SCs in 3D) and the amount of proteins secreted from two culturing systems were compared. The results show that the metabolic activity and the amount of secreted proteins from SCs(3D) were higher than both from SCs(2D). Three coculturing groups: NSCs+SC(2D), NSCs+SC(3D), and NSCs +SC-conditioned medium (SCCM, control group) were also compared regarding cell morphology and the numbers of neurons, neural outgrowths and neurospheres. The quantity of neurons, neural outgrowths and neurospheres were the highest in the NSCs+SC(3D) group. SCs cultured in the 3D system had a strong trophic effect on NSCs and enhanced their survival and growth. Besides, the mRNA of trophic and nutritive factors such as Glial-cell-line-derived neurotrophic factor (GDNF) and Interleukin-1 α (IL-1 α) secreted by the SCs from both 2D and 3D culture system were analyzed by real time-PCR and gel assay. The mRNA transcription of GDNF and IL-1α is more apparent in the 3D culture system than that from the 2D one. The coculturing system of NSCs+SC(3D) is a promising candidate for future neural stem cell transplantation.

Performance and Mechanism of Neuroleukin in the Growth and Survival of Sertoli Cell-Induced Neurons in a Coculture System

“…In previous study, we reported that SCs can enhance the growth and survival of NSCs and neurons (33,34). In this study, a transwell coculture system was employed in order to determine the mechanism: whether the promotion of neurite outgrowth and prosurvival were mediated only by putative secreted factors or regulated by cell surface-anchored factors.…”

“…In previous studies, we reported that expanded SCs could effectively stimulate outgrowth of NSCs in a coculture system (33,34). In this study, we showed that SCs promoted the survival and neurite outgrowth of neurons without direct contact.…”

Cell‐penetrating peptide–labelled smart polymers for enhanced gene delivery

“…Statistical analysis of the numerical variables was performed using a two‐sample, two‐tailed t ‐test. A value of p < 0.05 is considered to be significant .…”

Emerging Strategies for Stem Cell Lineage Commitment in Tissue Engineering and Regenerative Medicine

“…Although these signals can be presented to stem cells in vitro, such as by immobilization to materials, the large number of different signals and varying times for exposure may not allow for recapitulation of stem cell differentiation. In view of this, the presence of other cells in coculture with stem cells can be a useful tool to guide in vitro stem cell differentiation and lineage commitment. − …”