Modeling neural differentiation on micropatterned substrates coated with neural matrix components

García-Parra, Patricia; Cavaliere, Fabio; Maroto, Marcos; Bilbao, Leire; Obieta, Isabel; Munáin, Adolfo López de; Alava, J.I.; Izeta, Ander

doi:10.3389/fncel.2012.00010

Cited by 21 publications

(21 citation statements)

References 75 publications

(85 reference statements)

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“…Neural extracellular matrix (nECM)-coated substrates were developed as described [14]. Briefly, 12 mm-diameter coverslips were incubated with a solution of Cultrex® basement membrane extract (BME) without phenol red (8.32 mg/ml; Trevigen, Gaithersburg, USA; it is a soluble form of basement membrane purified from Engelbreth-Holm-Swarm tumor; the major components of this BME include laminin, collagen IV, entactin, and heparin sulfate proteoglycan), hyaluronan of low molecular weight (31 kDa, HLMW) (7.5 mg/ml; R&D, Minneapolis, USA), Netrin-G1a and Netrin-4 (25 μg/ml; R&D, Minneapolis, USA) in phosphate-buffered saline (PBS) (Life Technologies Corporation, Paisley, UK).…”

Section: Methodsmentioning

confidence: 99%

“…In a previous article we described the development, characterization and functional validation of a nECM-based polymeric support used as a biocompatible and efficient in vitro microenvironment for neuronal differentiation of both PC12 cells and adult skin-derived precursor cells [14]. Since nECM proteoglycans are involved in axonal regeneration and in post-traumatic neuronal plasticity in the CNS [15,16], the rationale for the present methodological article was to use the nECM component of this polymeric support to study neuronal development and, in particular, dopaminergic differentiation.…”

Section: Introductionmentioning

confidence: 99%

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A neural extracellular matrix-based method for in vitrohippocampal neuron culture and dopaminergic differentiation of neural stem cells

et al. 2013

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BackgroundThe ability to recreate an optimal cellular microenvironment is critical to understand neuronal behavior and functionality in vitro. An organized neural extracellular matrix (nECM) promotes neural cell adhesion, proliferation and differentiation. Here, we expanded previous observations on the ability of nECM to support in vitro neuronal differentiation, with the following goals: (i) to recreate complex neuronal networks of embryonic rat hippocampal cells, and (ii) to achieve improved levels of dopaminergic differentiation of subventricular zone (SVZ) neural progenitor cells.MethodsHippocampal cells from E18 rat embryos were seeded on PLL- and nECM-coated substrates. Neurosphere cultures were prepared from the SVZ of P4-P7 rat pups, and differentiation of neurospheres assayed on PLL- and nECM-coated substrates.ResultsWhen seeded on nECM-coated substrates, both hippocampal cells and SVZ progenitor cells showed neural expression patterns that were similar to their poly-L-lysine-seeded counterparts. However, nECM-based cultures of both hippocampal neurons and SVZ progenitor cells could be maintained for longer times as compared to poly-L-lysine-based cultures. As a result, nECM-based cultures gave rise to a more branched neurite arborization of hippocampal neurons. Interestingly, the prolonged differentiation time of SVZ progenitor cells in nECM allowed us to obtain a purer population of dopaminergic neurons.ConclusionsWe conclude that nECM-based coating is an efficient substrate to culture neural cells at different stages of differentiation. In addition, neural ECM-coated substrates increased neuronal survival and neuronal differentiation efficiency as compared to cationic polymers such as poly-L-lysine.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A neural extracellular matrix-based method for in vitrohippocampal neuron culture and dopaminergic differentiation of neural stem cells

et al. 2013

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“…Other soluble factors (i.e., biomolecules) are also difficult to focus spatially. Microfluidic devices and electrode arrays provide strategies to operate some stimulus exposure events on a moderately local scale [103,104]. High spatial resolution of stimulus application can be achieved with light [95].…”

Section: Stimulimentioning

confidence: 99%

Responsive Cell–Material Interfaces

Dhowre

Rajput

Russell

et al. 2015

Nanomedicine (Lond.)

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Major design aspects for novel biomaterials are driven by the desire to mimic more varied and complex properties of a natural cellular environment with man-made materials. The development of stimulus responsive materials makes considerable contributions to the effort to incorporate dynamic and reversible elements into a biomaterial. This is particularly challenging for cell-material interactions that occur at an interface (biointerfaces); however, the design of responsive biointerfaces also presents opportunities in a variety of applications in biomedical research and regenerative medicine. This review will identify the requirements imposed on a responsive biointerface and use recent examples to demonstrate how some of these requirements have been met. Finally, the next steps in the development of more complex biomaterial interfaces, including multiple stimuli-responsive surfaces, surfaces of 3D objects and interactive biointerfaces will be discussed.

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“…For example, laminin has been identified as the optimal substrate for long-term culture of NPCs (57). Along similar lines, a combination of collagen, laminin, entactin, and proteoglycans was developed as a neuronal ECM to support in vitro neural differentiation of NPCs (63,64). Natural ECMs such as fibronectin and laminin have been used as cell culture coatings for the continued proliferation of primary muscle stem cells in vitro (163).…”

Section: Natural Ecms Recapitulate In Vivo Biological Activitymentioning

confidence: 99%

Constructing stem cell microenvironments using bioengineering approaches

Brafman

2013

Physiological Genomics

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Within the adult organism, stem cells reside in defined anatomical microenvironments called niches. These architecturally diverse microenvironments serve to balance stem cell self-renewal and differentiation. Proper regulation of this balance is instrumental to tissue repair and homeostasis, and any imbalance can potentially lead to diseases such as cancer. Within each of these microenvironments, a myriad of chemical and physical stimuli interact in a complex (synergistic or antagonistic) manner to tightly regulate stem cell fate. The in vitro replication of these in vivo microenvironments will be necessary for the application of stem cells for disease modeling, drug discovery, and regenerative medicine purposes. However, traditional reductionist approaches have only led to the generation of cell culture methods that poorly recapitulate the in vivo microenvironment. To that end, novel engineering and systems biology approaches have allowed for the investigation of the biological and mechanical stimuli that govern stem cell fate. In this review, the application of these technologies for the dissection of stem cell microenvironments will be analyzed. Moreover, the use of these engineering approaches to construct in vitro stem cell microenvironments that precisely control stem cell fate and function will be reviewed. Finally, the emerging trend of using high-throughput, combinatorial methods for the stepwise engineering of stem cell microenvironments will be explored.

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Modeling neural differentiation on micropatterned substrates coated with neural matrix components

Cited by 21 publications

References 75 publications

A neural extracellular matrix-based method for in vitrohippocampal neuron culture and dopaminergic differentiation of neural stem cells

A neural extracellular matrix-based method for in vitrohippocampal neuron culture and dopaminergic differentiation of neural stem cells

Responsive Cell–Material Interfaces

Constructing stem cell microenvironments using bioengineering approaches

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