The olfactory bulb and olfactory mucosa obtained from human cadaver donors as a source of olfactory ensheathing cells

Abstract

During the last decade, olfactory ensheathing cells (OECs) have been successfully applied in multiple experimental approaches aimed to repair damaged mammalian spinal cord. Some of these experiments have consequently been translated into clinical trials. Finding a reliable source of human OECs that is easily accessible and can ensure a sufficient number of cells is a major prerequisite for conducting studies on OEC-mediated spinal cord regeneration. Here, we present a procedure for obtaining olfactory bulbs (OBs) and olfactory mucosa (OM) simultaneously from adult cadaver heart-beating donors for OEC isolation and analyze some of the factors that may condition successful OEC culture. We show that the results of OEC culture from OBs (10 cases) correlated significantly with warm ischemia time (WIT) as well as the initial viability of the isolated cells. Efficient OEC culture was possible when the WIT for the OB was up to 20 min. Brain damage, assessed by determination of S100B serum level, was not related to the success of OEC culture from the OB. Cadaver OM (7 cases) was shown to be a more reliable source of human OECs than the OB. In most of the examined cases the efficacy of culturing OECs from cadaver OM obtained even 180 min after cardiac arrest was comparable to that of living patients. The method of obtaining OBs and OM from cadavers enables the use of an alternative source of primary adult human OECs for further preclinical and clinical studies on their neurotrophic properties.

The Impact of Oxidative Stress Factors on the Viability, Senescence, and Methylation Status of Olfactory Bulb-Derived Glial Cells Isolated from Human Cadaver Donors

“…OB-derived glial cells have been intensively studied because of their neuroregenerative potential [Gudiño-Cabrera and Nieto-Sampedro, 1996;Międzybrodzki et al, 2006;Mackay-Sim et al, 2008;Chhabra et al, 2009, Grzesiak et al, 2015. Several research groups has pre- vealing a decreased viability of cells in the ≥20-h group.…”

“…Glial cells derived from OBs were isolated using modified procedures described previously [Międzybrodzki et al, 2006]. First, isolated OBs were placed into sterile Hanks' balanced salt solution (HBSS) containing 1% of antibiotic/solution (P/S/A; penicillin/ streptomycin/amphotericin B).…”

“…The duration of ischemia plays a crucial role in obtaining viable cells from postmortem tissues [Hwang et al, 2005;Międzybrodzki et al, 2006;Halazun et al, 2007]. The pathological mechanism of ischemia involves inflammation, glial inactivation, and programmed cell death.…”

“…Międzybrodzki et al [2006] presented a procedure for isolating OBs from adult cadaver donors and analyzed the viability of the OECs in culture. They concluded that warm ischemia time significantly affected the behavior and characteristics of the isolated cells.…”

“…In this study, we evaluated the characteristics of primary glial cell cultures derived from OBs isolated before and after 20 h after the death of the donor. The choice of the time period for the OB dissection was derived from earlier studies by Międzybrodzki et al [2006] and Gudiño-Cabrera and Nieto-Sampedro [1996].…”