The Immunohistochemical Characterization of Human Fetal Olfactory Bulb and Olfactory Ensheathing Cells in Culture as a Source for Clinical CNS Restoration

Abstract

Clinical studies have expanded the therapeutic olfactory ensheathing cells (OECs) transplantation to different human Central Nervous System (CNS) diseases. In fact, the OEC transplantation in clinic is a mixture of olfactory bulb cells; they even have not demonstrated that they have such a subpopulation yet. However, as a source of OECs transplantation, the development and identification of human fetal OECs are still need more understanding, because some surgery try to restoration CNS injury with a more purity of OEC cultures generated by a number of different procedures. In this article, twelve human fetal olfactory bulb (OB) samples were obtained from six fetuses in 20 weeks of gestation, it was studied by immunofluorescence on histological sections and cultured cells with multiple antibodies under confocal microscopy. The P75NTR positive OB-OECs (olfactory ensheathing cell from the olfactory bulb) were present in both outer olfactory nerve layers and glomerular layer. The percentage of OB cells in culture, about 22.31 was P75NTR positive, 45.77 was S100b, and 31.92 was GFAP. P75NTR and GFAP were coexpressed with S100b, respectively; however, P75NTR was not coexpressed with GFAP in human fetal OECs. It is suggested that the localization and development of human OECs in OB are different to those in rodent, and the P75NTR immunohistological staining is still necessary to identify and characterize human fetal OECs in culture before transplantation. Anat Rec, 293:359-369, 2010. V V C 2009 Wiley-Liss, Inc.

Common olfactory ensheathing glial markers in the developing human olfactory system

“…These results are similar to those found in adult canine, guinea pig and cat, but in contrast to rat and murine OECs (Gong et al 1994; Franceschini and Barnett 1996; Au et al 2002; Au and Roskams 2003; Bock et al 2007; Smithson and Kawaja 2009). These results are also in contrast with Liu et al (2010) who observed substantial P75 NTR immunoreactivity in the ONL and surrounding glomeruli in the olfactory bulb in 20 pcw human foetuses (Liu et al 2010). Differences between our study and Liu et al could be due to differing antibodies or foetal sample age.…”

“…No selective purification methods have been described to isolate OECs from other cells of the olfactory bulb indicating these cultures are likely to contain a wide range of cells and not just OECs. Liu et al (2010) characterised human foetal OB cultures generated using similar methods and found them to contain just 23 % P75 NTR + cells. Other cells in the cultures were GFAP+, S100+ and nestin+, possibly neural stem and precursor cells, reactive astrocytes or microglia (Liu et al 2010).…”

“…Liu et al (2010) characterised human foetal OB cultures generated using similar methods and found them to contain just 23 % P75 NTR + cells. Other cells in the cultures were GFAP+, S100+ and nestin+, possibly neural stem and precursor cells, reactive astrocytes or microglia (Liu et al 2010). Another report by Guest et al (2006) found similar cultures described as OECs contained nestin and GFAP but no S100 and were morphologically reminiscent of human neurosphere cultures; the transplants were not clearly OECs (Guest et al 2006).…”

A Neuroregenerative Human Ensheathing Glia Cell Line with Conditional Rapid Growth

“…Briefly, we used a previously described method (41) with modifications (31) to obtain OEG primary cultures from adult human olfactory bulbs that were obtained from the Tissue Bank for Neurological Research of Madrid. OEG were identified in the olfactory bulb following established morphological and anatomical guidelines (34) and cultured in ME medium: DMEM/F12 (1:1), 10% FCS, 2 mM glutamine, 20 μg/ml pituitary extract, 2 μM forskolin, 50 μg/ml primocin. Our previous study (31) also describes the infection of these human OEG with lentivectors encoding BMI1 and TERT, isolation of immortalized clones, and the characterization of the clonal lines as well as the primary cells for the expression of glial antigenic markers including S100β, GFAP, neuroligin-3, APP, vimentin, and nestin.…”

Olfactory ensheathing cells from the nasal mucosa and olfactory bulb have distinct membrane properties

“…Much of our current understanding of neuroglial interactions comes from structural, molecular, and functional studies of rodent OECs, from either the OM or OB (Guerout et al, 2010;Hayat, Wigley, & Robbins, 2003;Higashi et al, 2001;Piantanida et al, 2019;Rash et al, 2005;Rela, Bordey, & Greer, 2010;Rela, Piantanida, Bordey, & Greer, 2015;Rieger, Deitmer, & Lohr, 2007). Studies of excised human OM tissues have also helped identify common markers in vivo (Choi, Law, Raisman, & Li, 2008;Liu et al, 2010;Oprych, Cotfas, & Choi, 2017).…”