Emergence of oligodendrocytes from human neural spheres

Abstract

To study the development of human oligodendrocyte precursors (OP), we expanded human embryonic brain-derived neural precursors into spheres with basic fibroblast growth factor (FGF2). Over 90% of the cells in the expanded spheres were precursors coexpressing nestin and the polysialylated (PSA) form of NCAM. The remaining cells were mostly astrocytes and neuronal cells located at the periphery of the floating spheres. When spheres were allowed to adhere on fibronectin-coated substrate in the absence of FGF2, neural precursors migrated in the outgrowth and often formed chains of cells expressing high levels of PSA-NCAM. Many migrating cells also expressed beta-3 tubulin while only scattered elongated cells radiating from the spheres were GFAP+ astrocytes. Spindle-shaped cells not associated with the chains were labeled for the PDGF-alpha receptor and often coexpressed MAP2 neuronal isoforms. Neuronal cells in the outgrowth rapidly established a rich neuritic network where OP expressing O4 and DM20/proteolipid antigens appeared. T3 treatment of neural spheres increased the rate of OP formation and the complexity of their shape. Thus, the generation of human oligodendrocytes from neural precursors is tightly correlated with growth of neuronal processes and enhanced by hormonal signals.

Isolation of mineralizing Nestin+ Nkx6.1+ vascular muscular cells from the adult human spinal cord

“…In these conditions, we could not detect staining of GFP-labelled cells (100 individual cells examined) with neuronal (Map2ab), astrocytic (GFAP) or oligodendrocytic (Olig2, Nkx2.2) markers, strongly indicating the absence of neural differentiation of these cells (Figure 5). Third, we treated the cells with BMP2, BMP4, BMP7, CNTF, CNTF/BMP4, LIF, LIF/BMP4 or T3 to promote an astrocytic [22,23] or an oligodendrocytic fate [24] respectively, but again no differentiation was observed (data not shown).…”

Scientific Validation of Human Neurosphere Assays for Developmental Neurotoxicity Evaluation

“…In order to develop into myelinating OLs, OPCs and pre-OLs have to mature and express myelin-associated genes including myelin basic protein (MBP) and myelin proteolipid protein (PLP1). This maturation processes and the proper development of white matter tracts in humans depend on thyroid hormones (TH), such as the thyroxine metabolite triiodothyronine (T3; Annunziata et al, 1983 ; Baas et al, 1997 ; Murray and Dubois-Dalcq, 1997 ). In line with that, in vivo studies on hypothyroid rats reported reduced numbers of mature OLs and impaired expression of PLP1 and MBP ( Ibarrola and Rodríguez-Peña, 1997 ; Schoonover et al, 2004 ).…”

Bioelectric Potential in Next-Generation Organoids: Electrical Stimulation to Enhance 3D Structures of the Central Nervous System

“…However, to produce alternative CNS-derivative cells from PSCs, suitable biochemical cues were required. For example, culturing in the presence of epidermal growth factor (EGF) and fibroblast growth factor (FGF)-2, known to promote proliferation of glial precursor cells, and timed addition of tri-iodothyronine (T3), which induces oligodendrocytes, was used to derive glial cell lineages ( Murray and Dubois-Dalcq, 1997 ; Brüstle et al, 1999 ; Reubinoff et al, 2001 ).…”