A novel primary culture method for high-purity satellite glial cells derived from rat dorsal root ganglion

Wang, Xianbin; Ma, Wei; Luo, Tao; Yang, Jinwei; Wang, Xiangpeng; Dai, Yunfei; Guo, Jun; Li, Liyan

doi:10.4103/1673-5374.244797

Cited by 21 publications

(12 citation statements)

References 31 publications

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“…Several studies have characterized GS as an SGC‐specific marker in murine and rat sensory ganglia that identifies SGCs in situ and in vitro . 6 , 9 , 35 , 36 , 37 , 38 , 39 , 40 , 41 GS catalyses the conversion of the excitatory neurotransmitter glutamate to glutamine 42 and is expressed by SGCs together with glutamate receptors and glutamate transporters. 43 In the CNS, astrocytes represent the main glial cell population expressing GS.…”

Section: Discussionmentioning

confidence: 99%

Phenotypical peculiarities and species‐specific differences of canine and murine satellite glial cells of spinal ganglia

Huang

Zdora

Buhr

et al. 2021

J Cellular Molecular Medi

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Satellite glial cells (SGCs) of the sensory ganglia are a glial cell population of the peripheral nervous system (PNS) with diverse and remarkable features. Within the past two decades, SGCs have received increasing attention in a wide field of research. In addition to studying their morphological and functional qualities, their role in pathologic states and the development of neuropathic pain has been investigated. 1 The sensory ganglia, including the trigeminal ganglion as well as the spinal ganglia (SG), also known as dorsal root ganglia (DRG), are part of the PNS and transmit sensory signals fromthe periphery towards the central nervous system (CNS).

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Section: Discussionmentioning

confidence: 99%

Phenotypical peculiarities and species‐specific differences of canine and murine satellite glial cells of spinal ganglia

Huang

Zdora

Buhr

et al. 2021

J Cellular Molecular Medi

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“…Our obtained results are in accordance with the previously published data because only glial cells migrated from DRGs in 2D cultures. However, no outgrowing neurites could be detected [23].…”

Section: Comparison Of 2d and 3d Drg Explant Culture Modelsmentioning

confidence: 93%

Three-Dimensional Model of Dorsal Root Ganglion Explant as a Method of Studying Neurotrophic Factors in Regenerative Medicine

et al. 2020

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Neurotrophic factors play a key role in the development, differentiation, and survival of neurons and nerve regeneration. In the present study, we evaluated the effect of certain neurotrophic factors (NGF, BDNF, and GDNF) on axon growth and migration of Nestin-green fluorescent protein (GFP)-positive cells using a 3D model of dorsal root ganglion (DRG) explant culture in Matrigel. Our method generally represents a convenient model for assessing the effects of soluble factors and therapeutic agents on axon growth and nerve regeneration in R&D studies. By analyzing the DRG explants in ex vivo culture for 21 days, one can evaluate the parameters of neurite outgrowth and the rate of cell migration from the DRG explants into the Matrigel. For the current study, we used Nestin-GFP-expressing mice in which neural precursors express Nestin and the green fluorescent protein (GFP) under the same promoter. We revealed that GDNF significantly (two fold) stimulated axon outgrowth (p < 0.05), but not BDNF or NGF. It is well-known that axon growth can be stimulated by activated glial cells that fulfill a trophic function for regenerating nerves. For this reason, we evaluated the number of Nestin-GFP-positive cells that migrated from the DRG into the Matrigel in our 3D ex vivo explant model. We found that NGF and GDNF, but not BDNF, stimulated the migration of Nestin-GFP cells compared to the control (p < 0.05). On the basis of the aforementioned finding, we concluded that GDNF had the greatest stimulating potential for axon regeneration, as it stimulated not only the axon outgrowth, but also glial cell migration. Although NGF significantly stimulated glial cell migration, its effect on axon growth was insufficient for axon regeneration.Biomedicines 2020, 8, 49 2 of 13 (ECM) that surrounds the cells and creates the microenvironment in vivo [6,8]. The lack of the proper cell-cell and cell-matrix contacts results in change of gene and protein expression profiles [9]. These can adversely affect cell morphology, cell cycle progression, cell survival, and intracellular signaling, potentially resulting in false positive or false negative data acquisition [8,10].In 2D culturing systems, the bioavailability of substances is difficult to evaluate [11]. All cells in a monolayer in 2D culturing conditions simultaneously receive the same dose of a tested agent, whereas in vivo there is a gradient of a tested agent that penetrates the matrix and several layers of cells in a tissue [12]. 3D cultures allow for the assessment of the parameters of a gradual diffusion of tested agents through the gel [6,11].Primary neural cell cultures represent a modern challenge in cell biology, as mature neurons do not undergo cell division in vitro, making it complicated to obtain a sufficient number of cells for experiments [13]. Cell survival is usually low, and neural cell cultures from DRG can be maintained for only 5-7 days, provided that soluble factors that increase survival are added [14]. This is one of the complications in result interpretation when...

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“…SGCs were identified by the expression of SGC markers S100B and Fabp7 (Figure 1a; Figure S1) (Wang et al, 2019;Weider et al, 2015).…”

Section: Identification Of Cell Clusters and Marker Genesmentioning

confidence: 99%

Single‐cell transcriptomic profiling of satellite glial cells in stellate ganglia reveals developmental and functional axial dynamics

Weperen

Littman²,

Arneson

et al. 2021

Glia

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Stellate ganglion neurons, important mediators of cardiopulmonary neurotransmission, are surrounded by satellite glial cells (SGCs), which are essential for the function, maintenance, and development of neurons. However, it remains unknown whether SGCs in adult sympathetic ganglia exhibit any functional diversity, and what role this plays in modulating neurotransmission. We performed single‐cell RNA sequencing of mouse stellate ganglia (n = 8 animals), focusing on SGCs (n = 11,595 cells). SGCs were identified by high expression of glial‐specific transcripts, S100b and Fabp7. Microglia and Schwann cells were identified by expression of C1qa/C1qb/C1qc and Ncmap/Drp2, respectively, and excluded from further analysis. Dimensionality reduction and clustering of SGCs revealed six distinct transcriptomic subtypes, one of which was characterized the expression of pro‐inflammatory markers and excluded from further analyses. The transcriptomic profiles and corresponding biochemical pathways of the remaining subtypes were analyzed and compared with published astrocytic transcriptomes. This revealed gradual shifts of developmental and functional pathways across the subtypes, originating from an immature and pluripotent subpopulation into two mature populations of SGCs, characterized by upregulated functional pathways such as cholesterol metabolism. As SGCs aged, these functional pathways were downregulated while genes and pathways associated with cellular stress responses were upregulated. These findings were confirmed and furthered by an unbiased pseudo‐time analysis, which revealed two distinct trajectories involving the five subtypes that were studied. These findings demonstrate that SGCs in mouse stellate ganglia exhibit transcriptomic heterogeneity along maturation or differentiation axes. These subpopulations and their unique biochemical properties suggest dynamic physiological adaptations that modulate neuronal function.

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A novel primary culture method for high-purity satellite glial cells derived from rat dorsal root ganglion

Cited by 21 publications

References 31 publications

Phenotypical peculiarities and species‐specific differences of canine and murine satellite glial cells of spinal ganglia

Phenotypical peculiarities and species‐specific differences of canine and murine satellite glial cells of spinal ganglia

Three-Dimensional Model of Dorsal Root Ganglion Explant as a Method of Studying Neurotrophic Factors in Regenerative Medicine

Single‐cell transcriptomic profiling of satellite glial cells in stellate ganglia reveals developmental and functional axial dynamics

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