Xeno‐free culture for generation of forebrain oligodendrocyte precursor cells from human pluripotent stem cells

Abstract

Oligodendrocytes (OLs) show heterogeneous properties that depend on their location in the central nervous system (CNS). In this regard, the investigation of oligodendrocyte precursor cells (OPCs) derived from human pluripotent stem cells (hPSCs) should be reconsidered, particularly in cases of brain‐predominant disorders for which brain‐derived OPCs are more appropriate than spinal cord‐derived OPCs. Furthermore, animal‐derived components are responsible for culture variability in the derivation and complicate clinical translation. In the present study, we established a xeno‐free system to induce forebrain OPCs from hPSCs. We induced human forebrain neural stem cells (NSCs) on Laminin 511‐E8 and directed the differentiation to the developmental pathway for forebrain OLs with SHH and FGF signaling. OPCs were characterized by the expression of OLIG2, NKX2.2, SOX10, and PDGFRA, and subsequent maturation into O4+ cells. In vitro characterization showed that >85% of the forebrain OPCs (O4+) underwent maturation into OLs (MBP+) 3 weeks after mitogen removal. Upon intracranial transplantation, the OPCs survived, dispersed in the corpus callosum, and matured into (GSTπ+) OLs in the host brains 3 months after transplantation. These findings suggest our xeno‐free induction of forebrain OPCs from hPSCs could accelerate clinical translation for brain‐specific disorders.

Sonic Hedgehog Pathway Modulation Normalizes Expression of Olig2 in Rostrally Patterned NPCs With Trisomy 21

“…The cells were differentiated to NPCs and transitioned towards an OL fate following a published protocol with some modifications (Douvaras and Fossati, 2015 ; Figure 1A ). PAX6, a classical marker for NPCs, was used to confirm the presence of NPCs at culture day 8 programmed either with Wnt-C59 (a Wnt pathway inhibitor, a driver of rostral CNS fate; Patapoutian and Reichardt, 2000 ; Hermanto et al, 2019 ) or Retinoic Acid (RA, a driver of caudal CNS fate; Lara-Ramírez et al, 2013 ; Goldman and Kuypers, 2015 ; Figures 1F–H ). Confirming the efficacy of Wnt-C59, we found reduced mRNA expression of Axin2, a downstream target of the Wnt pathway (Jho et al, 2002 ), compared to cells cultured with RA (WC eu 0.26x p = 0.0027, WC ts 0.46x p = 0.0023; ILD eu 0.45x p = 0.066, ILD ts 0.26x p = 0.0015; Figure 1B ).…”

Sonic Hedgehog Pathway Modulation Normalizes Expression of Olig2 in Rostrally Patterned NPCs with Trisomy 21

“…The cells were differentiated to NPCs and transitioned towards an OL fate following a published protocol with some modifications (Douvaras and Fossati, 2015) (Figure 1A). PAX6, a classical marker for NPCs, was used to confirm the presence of NPCs at culture day 8 programmed either with Wnt-C59 (a Wnt pathway inhibitor, a driver of rostral CNS fate (Hermanto et al, 2019;Patapoutian and Reichardt, 2000)) or Retinoic Acid (RA, a driver of caudal CNS fate (Goldman and Kuypers, 2015;Lara-Ramírez et al, 2013)) (Figure 1F,G,H).…”

The Role of Stem Cells in the Therapy of Stroke

“…Tissue survival was associated with decreases in inflammatory markers, glial scarring, and apoptotic neurons at both mRNA and protein levels. One of the transplant's therapeutic mechanisms is to replace damaged neurons, which involves the ability of the implanted cells to migrate, survive, proliferate, and differentiate into different types of cells that are needed [ 133 ]. However, the implantation of cells is limited and some of the cells remain undifferentiated.…”