Long-term tripotent differentiation capacity of human neural stem (NS) cells in adherent culture

Sun, Yirui; Pollard, Steven M.; Conti, Luciano; Toselli, Mauro; Biella, Gerardo; Parkin, Georgina; Willatt, Lionel; Falk, Anna; Cattaneo, Elena; Smith, Austin

doi:10.1016/j.mcn.2008.02.014

Cited by 206 publications

(231 citation statements)

References 44 publications

Supporting

Mentioning

218

Contrasting

Order By: Relevance

“…hsp-NSCs and hiPS-lt-NES cells were established and maintained as described previously [10,11,16]. The hsp-NS cell line CB660sp and hiPS-lt-NES cell line AF22 were used in the present study.…”

Section: Cell Culturementioning

confidence: 99%

Treatment of a Mouse Model of Spinal Cord Injury by Transplantation of Human Induced Pluripotent Stem Cell-Derived Long-Term Self-Renewing Neuroepithelial-Like Stem Cells

et al. 2012

Self Cite

View full text Add to dashboard Cite

Because of their ability to self-renew, to differentiate into multiple lineages, and to migrate toward a damaged site, neural stem cells (NSCs), which can be derived from various sources such as fetal tissues and embryonic stem cells, are currently considered to be promising components of cell replacement strategies aimed at treating injuries of the central nervous system, including the spinal cord. Despite their efficiency in promoting functional recovery, these NSCs are not homogeneous and possess variable characteristics depending on their derivation protocols. The advent of induced pluripotent stem (iPS) cells has provided new prospects for regenerative medicine. We used a recently developed robust and stable protocol for the generation of long-term, self-renewing, neuroepithelial-like stem cells from human iPS cells (hiPS-lt-NES cells), which can provide a homogeneous and well-defined population of NSCs for standardized analysis. Here, we show that transplanted hiPS-lt-NES cells differentiate into neural lineages in the mouse model of spinal cord injury (SCI) and promote functional recovery of hind limb motor function. Furthermore, using two different neuronal tracers and ablation of the transplanted cells, we revealed that transplanted hiPS-lt-NES cell-derived neurons, together with the surviving endogenous neurons, contributed to restored motor function. Both types of neurons reconstructed the corticospinal tract by forming synaptic connections and integrating neuronal circuits. Our findings indicate that hiPS-lt-NES transplantation represents a promising avenue for effective cell-based treatment of SCI. Disclosure of potential conflicts of interest is found at the end of this article.

show abstract

Section: Cell Culturementioning

confidence: 99%

Treatment of a Mouse Model of Spinal Cord Injury by Transplantation of Human Induced Pluripotent Stem Cell-Derived Long-Term Self-Renewing Neuroepithelial-Like Stem Cells

et al. 2012

Self Cite

View full text Add to dashboard Cite

show abstract

“…NSCs can be isolated from the mammalian central nervous system (CNS) and can be maintained in vitro for moderate periods of time without loss of the ability of these cells to be passaged (self-renew) and to differentiate into neurons, astrocytes, and oligodendrocytes, the primary cell types in the CNS (Dhara and Stice 2008;Sun et al 2008). A major challenge, however, is the expansion to obtain sufficient NSCs for these applications.…”

Section: Introductionmentioning

confidence: 99%

Optimal time for passaging neurospheres based on primary neural stem cell cultures

et al. 2011

View full text Add to dashboard Cite

Cultured neural stem cells (NSCs) provide a powerful means for investigating central nervous system disease, neuron development, differentiation, and regeneration. To obtain sufficient neurospheres, subculturing is essential following establishment of the primary NSC culture. Passaging the primary neurospheres is a key issue that is often ignored. We evaluated the influence of different passaging schedules on primary cultured NSCs. Passaging was performed on day 5, 7 or 9. We observed more neurospheres with diameters of 200-250 lm on day 7 than on day 5 or 9. Prolonging the time of primary culture reduced the cell metabolic activity by the MTT assay and cell proliferation by colony-forming assay and the differentiation to neurons from cells at P2 and later decreased. Additionally, more cells were in G0/G1 phase, and higher expression of p16INK4a and lower expression of cyclin D1 was found when the time of primary culture was prolonged to 9 days compared to 7-days cultures. Thus, in this study, we established that the optimal time for subculturing aggregated NSCs was on day 7 based on the primary culture.

show abstract

“…Furthermore, advances in stem cell research are opening new perspectives on the study and treatment of neurodegenerative disease, brain injury and brain cancer. Mouse and human neural stem (NS) cell lines can be maintained long term in vitro without genetic modification (Reynolds and Weiss, 1992;Svendsen et al, 1998;Carpenter et al, 1999;Riaz et al, 2002;Conti et al, 2005;Pollard et al, 2006;Sun et al, 2008). These cells offer a renewable resource for neurodegenerative disease studies and are suitable for pharmaceutical and neurotoxicological screening (Conti and Cattaneo, 2010).…”

Section: Introductionmentioning

confidence: 99%

“…We have previously reported the establishment of adherent mouse and human NS cell lines from various sources (Conti et al, 2005;Pollard et al, 2006;Sun et al, 2008). Monolayer culture enables mouse and human NS cells to retain stable self-renewal and clonal tripotent differentiation capacity over extended passaging without accompanying differentiation.…”

Section: Introductionmentioning

confidence: 99%

Interplay between FGF2 and BMP controls the self-renewal, dormancy and differentiation of rat neural stem cells

Sun

Zhou

et al. 2011

Development

Self Cite

View full text Add to dashboard Cite

Results Establishment of rat foetal NS cell culturesWe followed the protocol established for foetal mouse tissue described in Conti et al. (Conti et al., 2005). Dissociated cells from E13.5 rat foetal cortex or spinal cord were plated onto laminincoated tissue culture plastic in medium supplemented with FGF2 and epidermal growth factor (EGF). Many cells attached within 24 hours. The primary cultures were heterogeneous; they contained nestin-positive precursor cells, TuJ1-positive neurons and a small Summary Mouse and human central nervous system progenitor cells can be propagated extensively ex vivo as stem cell lines. For the rat, however, in vitro expansion has proven to be problematic owing to proliferation arrest and differentiation. Here, we analyse the establishment, in adherent culture, of undifferentiated tripotent neural stem (NS) cell lines derived from rat foetal brain and spinal cord. Rat NS cells invariably undergo growth arrest and apparent differentiation after several passages; however, conditioned medium from proliferating cultures can overcome this block, enabling continuous propagation of undifferentiated rat NS cells. We found that dormancy is induced by autocrine production of bone morphogenetic proteins (BMPs). Accordingly, the BMP antagonist noggin can replace conditioned medium to sustain continuous self-renewal. Noggin can also induce dormant cells to re-enter the cell cycle, upon which they reacquire neurogenic potential. We further show that fibroblast growth factor 2 (FGF2) is required to suppress terminal astrocytic differentiation and maintain stem cell potency during dormancy. These findings highlight an extrinsic regulatory network, comprising BMPs, BMP antagonists and FGF2 signals, that governs the proliferation, dormancy and differentiation of rat NS cells and which can be manipulated to enable long-term clonogenic self-renewal.

show abstract

Long-term tripotent differentiation capacity of human neural stem (NS) cells in adherent culture

Cited by 206 publications

References 44 publications

Treatment of a Mouse Model of Spinal Cord Injury by Transplantation of Human Induced Pluripotent Stem Cell-Derived Long-Term Self-Renewing Neuroepithelial-Like Stem Cells

Treatment of a Mouse Model of Spinal Cord Injury by Transplantation of Human Induced Pluripotent Stem Cell-Derived Long-Term Self-Renewing Neuroepithelial-Like Stem Cells

Optimal time for passaging neurospheres based on primary neural stem cell cultures

Interplay between FGF2 and BMP controls the self-renewal, dormancy and differentiation of rat neural stem cells

Contact Info

Product

Resources

About