Bilateral Interaction Between Cord Blood–Derived Human Neural Stem Cells and Organotypic Rat Hippocampal Culture

Abstract

The umbilical cord blood-derived neural stem/progenitor cells (HUCB-NSCs) potentially represent a rich source of transplantable material for treatment of a wide range of neurological diseases. Although, recently reported effects of their implementation in animal models of brain pathology are still controversial. As a simplified alternative to in vivo transplantation in this work we have applied a long-term organotypic rat hippocampal slice culture (OHC) as a recipient tissue to study bilateral graft/host cells interactions ex vivo. This type of culture can be considered as a kind of reductionistic model of brain transplantation where direct influence of systemic immunological responses to transplanted human cells would be excluded. The transplantation material derived from a HUCB-NSC line developed and characterized in our laboratory and delivered to the slices either as a single-cell suspension or after formation of typical neurospheres in serum-free medium in vitro (N-HUCBs). Experiments were focused on space-temporal context of cell transplantation in relation to their ability to ingrown, migrate, and differentiate within the slice cytoarchitecture. We gain evidences that these responses are strictly dependent on the engraftment site and that cell movement reflects typical routes used for migratory neuroblasts in vivo. The cells implanted at the second week of slice cultivation ingrown readily and deeply into host cytoarchitecture then matured to the level never observed in our transplantation animal models in vivo. Importantly, transplanted neurospheres, in addition to yield exogenous migratory cells to the host tissue can locally inhibit astrocytosis and promote outgrow of DCX-reactive neuroblasts in the surrounding OHC tissue.

Human Neural Progenitor Cells Show Functional Neuronal Differentiation and Regional Preference After Engraftment onto Hippocampal Slice Cultures

“…The finding does not appear to result from differences in the medium composition, as hNPCs growing on the insert membrane continued to express both markers. Interestingly, a similar suppression of GFAP expression was reported for human cord blood stem cells [12], whereas studies using mouse stem cells observed astrocytic differentiation in hippocampal cultures [9,10] and in striatal cultures [46], hinting at speciesspecific responses to the environment. The cause of this change and the extent to which the cells differ as a result, particularly in their signaling properties, require further investigation.…”

“…It has previously been demonstrated that when seeded onto organotypic hippocampal slice cultures, mouse embryonic stem cells can differentiate into functional neurons that receive synaptic input [8] and into glia [9,10]. Expression of neuronal markers has also been induced in mouse bone marrow stromal cells [11] and human cord blood-derived stem cells [12]; however, neural progenitor cells (NPCs) derived from adult rats predominantly differentiated into astrocytes when cocultured with hippocampal slices [13]. It is clear that the responsiveness to environmental cues varies between cell types, and as yet it is not clear how such environmental conditions might influence human cells.…”

Effects of Blast Overpressure on Neurons and Glial Cells in Rat Organotypic Hippocampal Slice Cultures

“…From 4 to 7 days in vitro (DIV), serum-based culture medium was gradually changed to a serum-free medium consisting of 50% MEM-Hank’s, 25% HBSS, 25% Neurobasal-A medium, 17 mM HEPES, 2 mM l -Glutamine, 2% B-27, 5 mg/ml d -glucose, and 1% antibiotic/antimycotic (all obtained from GIBCO Life Technologies). From 7 DIV until the end of the experiment, OHCs were grown in a serum-free medium ( 64 , 66 , 67 ) to decrease astrocyte proliferation and microglial activation ( 68 , 69 ). All PI uptake and LDH release measurements, described below in more detail, were also performed in the serum-free medium due to the interference of serum with these cell viability assays ( 70 , 71 ).…”

Experimental therapies for repair of the central nervous system: stem cells and tissue engineering

“…Transplantation of restricted neuronal progenitors is recognized by a number of authorities in the fields of neuroscience and regenerative medicine (Goldman, ; Hofstetter et al ., ; Lindvall et al ., ; Park et al ., ). This has been further confirmed by preclinical studies in which human neuroblasts generated from cord stem cells were transplanted into organotypic rat hyppocampal culture models (Sarnowska et al ., , ; Jurga et al ., ). The first human neuro‐implantation trial for stroke was reported in 2000 (Kondziolka et al ., ).…”