Embryonic stem cells produce neurotrophins in response to cerebral tissue extract: Cell line‐dependent differences

Abstract

In the present study, we compare the capacity of two different embryonic stem (ES) cell lines to secrete neurotrophins in response to cerebral tissue extract derived from healthy or injured rat brains. The intrinsic capacity of the embryonic cell lines BAC7 (feeder cell-dependent cultivation) to release brain-derived neurotrophic factor (BDNF) or neurotrophin-3 (NT-3) exceeded the release of these factors by CGR8 cells (feeder cell-free growth) by factors of 10 and 4, respectively. Nerve growth factor (NGF) was secreted only by BAC7 cells. Conditioning of cell lines with cerebral tissue extract derived from healthy or fluid percussion-injured rat brains resulted in a significant time-dependent increase in BDNF release in both cell lines. The increase in BDNF release by BAC7 cells was more pronounced when cells were incubated with brain extract derived from injured brain. However, differences in neurotrophin release associated with the origin of brain extract were at no time statistically significant. Neutrophin-3 and NGF release was inhibited when cell lines were exposed to cerebral tissue extract. The magnitude of the response to cerebral tissue extract was dependent on the intrinsic capacity of the cell lines to release neurotrophins. Our results clearly demonstrate significant variations in the intrinsic capability of different stem cell lines to produce neurotrophic factors. Furthermore, a significant modulation of neurotrophic factor release was observed following conditioning of cell lines with tissue extract derived from rat brains. A significant modulation of neurotrophin release dependent on the source of cerebral tissue extract used was not observed.

Extract Derived from Rat Brains in the Acute Phase Following Traumatic Brain Injury Impairs Survival of Undifferentiated Stem Cells and Induces Rapid Differentiation of Surviving Cells

“…However, a significant disadvantage to the use of ES cells has been the requirement to employ feeder layers of embryonic fibroblasts to support the isolation and maintenance of pluripotential stem cells. The stem cell line CGR8 has already been demonstrated to exhibit similar response pattern as a feeder cell dependent stem cell line [22]. The use of the stem cell line CGR8 thereby allows an unambiguous examination of the effect of the cerebral environment on differentiation in vitro as underlined by the findings reported in this paper.…”

“…Conditioning of different embryonic stem cell lines, including the cell line CGR8, with extract from healthy and injured rat brains resulted in a pronounced release of BDNF [22]. However, there was no significant difference in the amount of BDNF release when cells were incubated with TBE or HBE.…”

Human Embryonic Stem Cells (hESCs) Cultured Under Distinctive Feeder-Free Culture Conditions Display Global Gene Expression Patterns Similar to hESCs from Feeder-Dependent Culture Conditions

“…These advantages include minimal batch-tobatch variability, the high reproducibility of experimental outcomes, the easy standardization of experimental conditions, and the easy upscaling of the hESC cultures. Furthermore, the effects of genetic modifications and chemical treatments on the physiology of the hESCs would not be obscured by alterations in the feeder cell conditions, which are often observed in feeder-dependent hESC cultures [13].…”

Impurity of Stem Cell Graft by Murine Embryonic Fibroblasts – Implications for Cell-Based Therapy of the Central Nervous System

“…Various interactions between different cell types and the brain microenvironment were reported in several studies (Bentz et al, 2006, 2007, 2010; Molcanyi et al, 2007). We previously showed that incubation of ESCs with brain extract in vitro resulted in the release of neurotrophic factors, which was accompanied by the considerable co-production of these neurotrophins by inactivated co-cultured MEFs (Bentz et al, 2007). The metabolic potential of inactivated feeder cells is not surprising as MEFs are expected to produce humoral factors, which maintain the characteristics of co-cultured ESCs.…”