Maintaining multipotent trunk neural crest stem cells as self-renewing crestospheres

Mohlin, Sofie; Kunttas, Ezgi; Persson, C.; Abdel-Haq, Reem; Castillo, Aldo; Murko, Christina; Bronner, Marianne E.; Kerosuo, Laura

doi:10.1016/j.ydbio.2019.01.010

Cited by 16 publications

(22 citation statements)

References 40 publications

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“…Neural crest‐derived crestosphere cultures 24,25 enable studies on stemness properties of neural crest cells in vitro. Therefore, we examined EPAS1 expression in crestosphere cultures, in which multipotent neural crest cells can be maintained in a stem cell‐like state in vitro 25,26 .…”

Section: Resultsmentioning

confidence: 99%

“…For cranial‐derived cultures, the very anterior tip was excluded, and the neural tube was dissected until the first somite level as previously described 26 . For trunk‐derived cultures, the neural tube was dissected between somite 10 to 15 as previously described 24,25 . Pools of neural tubes from four to six embryos were used for each culture.…”

Section: Methodsmentioning

confidence: 99%

“…Single cells were transferred to 96-well plates prepared with 100 μL of NC medium supplemented with RA and BMP-4. 25 The absolute number of spheres formed in each well was quantified manually under the microscope. Sphere diameter was manually measured using the ImageJ software (spheres measured n = 33 and n = 27 for CTRL and EPAS1.2, respectively).…”

Section: Neural Tube Dissections For Crestosphere Culturesmentioning

confidence: 99%

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Hypoxia inducible factor‐2α importance for migration, proliferation, and self‐renewal of trunk neural crest cells

Niklasson

Fredlund

Monni

et al. 2020

Developmental Dynamics

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Background: The neural crest is a transient embryonic stem cell population. Hypoxia inducible factor (HIF)-2α is associated with neural crest stem cell appearance and aggressiveness in tumors. However, little is known about its role in normal neural crest development. Results: Here, we show that HIF-2α is expressed in trunk neural crest cells of human, murine, and avian embryos. Knockdown as well as overexpression of HIF-2α in vivo causes developmental delays, induces proliferation, and selfrenewal capacity of neural crest cells while decreasing the proportion of neural crest cells that migrate ventrally to sympathoadrenal sites. Reflecting the in vivo phenotype, transcriptome changes after loss of HIF-2α reveal enrichment of genes associated with cancer, invasion, epithelial-to-mesenchymal transition, and growth arrest. Conclusions: Taken together, these results suggest that expression levels of HIF-2α must be strictly controlled during normal trunk neural crest development and that dysregulated levels affects several important features connected to stemness, migration, and development. K E Y W O R D S embryogenesis, HIF-2α, migration, neural crest, stem cells, trunk neural crest Camilla U. Niklasson and Elina Fredlund contributed equally to this work.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Neural Tube Dissections For Crestosphere Culturesmentioning

confidence: 99%

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Hypoxia inducible factor‐2α importance for migration, proliferation, and self‐renewal of trunk neural crest cells

Niklasson

Fredlund

Monni

et al. 2020

Developmental Dynamics

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“…[82][83][84][85][86] Substantial amount of work has also been invested in revealing regulatory links between these transcription factors. 87,88 In 94 BMP4 has been identified as a pluripotency factor that promotes self-renewal in mouse embryonic stem cells (ESCs), 95 in human periodontal ligament cells 96 and adipose derived stem cells. 97 Although several other signaling molecules also undoubtedly contribute to the stem cell properties of NC cells, we are yet to uncover the mechanisms of action and the downstream and upstream players involved.…”

Section: How Is Stem Cell Potential Established and Maintained In Tmentioning

confidence: 99%

On the road again: Establishment and maintenance of stemness in the neural crest from embryo to adulthood

Perera

Kerosuo

2020

Stem Cells

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Unique to vertebrates, the neural crest (NC) is an embryonic stem cell population that contributes to a greatly expanding list of derivatives ranging from neurons and glia of the peripheral nervous system, facial cartilage and bone, pigment cells of the skin to secretory cells of the endocrine system. Here, we focus on what is specifically known about establishment and maintenance of NC stemness and ultimate fate commitment mechanisms, which could help explain its exceptionally high stem cell potential that exceeds the “rules set during gastrulation.” In fact, recent discoveries have shed light on the existence of NC cells that coexpress commonly accepted pluripotency factors like Nanog, Oct4/PouV, and Klf4. The coexpression of pluripotency factors together with the exceptional array of diverse NC derivatives encouraged us to propose a new term “pleistopotent” (Greek for abundant, a substantial amount) to be used to reflect the uniqueness of the NC as compared to other post-gastrulation stem cell populations in the vertebrate body, and to differentiate them from multipotent lineage restricted stem cells. We also discuss studies related to the maintenance of NC stemness within the challenging context of being a transient and thus a constantly changing population of stem cells without a permanent niche. The discovery of the stem cell potential of Schwann cell precursors as well as multiple adult NC-derived stem cell reservoirs during the past decade has greatly increased our understanding of how NC cells contribute to tissues formed after its initial migration stage in young embryos.

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“…With the spectacular development of the stem cell field, the last decade witnessed the establishment of several procedures to produce neural crest stem cells from embryonic, fetal, and adult tissues. Thus, the use of embryonic stem cells, induced pluripotent stem cells, neuroepithelial ''crestospheres'' that self-renew and retain multipotency for weeks, or reprogrammed cells from adult teeth, skin, or nerves proved to be extremely potent, in particular for generating large amounts of cells usable for comprehensive large scale analyses, such epigenomic and transcriptome profiling (Bajpai et al, 2017;Kerosuo, Nie, Bajpai, & Bronner, 2015;Liu & Cheung, 2016;Mohlin et al, 2019;Prescott et al, 2015). Mastering primary cultures of neural crest cells certainly provides the fundamentals and the knowledge for developing these sophisticated approaches.…”

Section: Commentary Background Informationmentioning

confidence: 99%

Establishing Primary Cultures of Trunk Neural Crest Cells

2020

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Neural crest cells constitute a unique population of progenitor cells with extensive stem cell capacities able to navigate throughout various environments in the embryo and are a source of multiple cell types, including neurons, glia, melanocytes, smooth muscles, endocrine cells, cardiac cells, and also skeletal and supportive tissues in the head. Neural crest cells are not restricted to the embryo but persist as well in adult tissues where they provide a reservoir of stem cells with great therapeutic promise. Many fundamental questions in cell, developmental, and stem cell biology can be addressed using this system. During the last decades there has been an increased availability of elaborated techniques, animal models, and molecular tools to tackle neural crest cell development. However, these approaches are often very challenging and difficult to establish and they are not adapted for rapid functional investigations of mechanisms driving cell migration and differentiation. In addition, they are not adequate for collecting pure populations of neural crest cells usable in large scale analyses and for stem cell studies. Transferring and adapting the neural crest system in tissue culture may then represent an attractive alternative, opening up numerous prospects. Here we describe a simple method for establishing primary cultures of neural crest cells derived from trunk neural tubes using the avian embryo as a source of cells. This protocol is suited for producing pure populations of neural crest cells that can be processed for cytological, cellular, and functional approaches aimed at characterizing their phenotype, behavior, and potential. © 2020 Wiley Periodicals LLC. Basic Protocol: Primary cultures of avian trunk neural crest cells Support Protocol: Adaptations for immunofluorescence labeling and videomicroscopy

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Maintaining multipotent trunk neural crest stem cells as self-renewing crestospheres

Cited by 16 publications

References 40 publications

Hypoxia inducible factor‐2α importance for migration, proliferation, and self‐renewal of trunk neural crest cells

Hypoxia inducible factor‐2α importance for migration, proliferation, and self‐renewal of trunk neural crest cells

On the road again: Establishment and maintenance of stemness in the neural crest from embryo to adulthood

Establishing Primary Cultures of Trunk Neural Crest Cells

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