Biofabrication of a three dimensional human‐based personalized neurofibroma model

Abstract

Neurofibromas are the most characteristic feature of neurofibromatosis type 1 (NF1), a multisystemic disorder caused by aberrations in the neurofibromin gene (NF1). Despite significant progress over the last several years in understanding this disease, a suitable in vitro model to better mimic neurofibroma formation and growth has yet to be described. There is therefore a need to establish an in vitro, three dimensional model that allows the incorporation of multicellular lineages and the modulation of the cellular microenvironment—known to be important for cellular crosstalk and distribution of soluble factors—to study neurofibroma biology and morphogenesis. A self‐assembly approach was used to generate tissue‐engineered skins (TES) in which patient‐derived spheroids made of NF1‐associated Schwann cells and fibroblasts were seeded. We describe the first in vitro three dimensional neurofibroma model—directly derived from NF1 patients presenting with histopathological features—having an ECM protein expression profile quite similar to that of a native tumor. We observed efficient incorporation, proliferation, and migration of spheroids within NF1‐TES over time. This biotechnological approach could provide a unique tool for precision medicine targeting NF1 and for assessing the tumorigenic properties of each NF1 gene mutation linked to tumor formation.

Spherical rotary cell seeding system for production of small-caliber tissue-engineered blood vessels with complex geometry

“…A more uniform cellular distribution may then help tissue maturation and consequently improve ECM secretion and assembly [32][33][34] . It has been shown that cell-cell and cell-ECM interactions in tissue engineering are needed in disease modeling to better represent crosstalk between cells, tissue compositions and the microenvironment associated to complex human pathologies 11,35,36 .…”

“…Macroscopic photographs of the biopsied TEBVs junction sites were also taken prior to the histological analysis. Fixed 10 μm TEBV cross-sections were then stained with hematoxylin and eosin (H&E) as previously described 36 . Microscopic images were acquired and measured under bright-field conditions using an upright microscope (AxioImager.M2; Carl Zeiss Microscopy, Jena, TH, Germany).…”

In Vitro Characterization of Motor Neurons and Purkinje Cells Differentiated from Induced Pluripotent Stem Cells Generated from Patients with Autosomal Recessive Spastic Ataxia of Charlevoix-Saguenay

“…We showed for the first time that it was possible to differentiate ARSACS-iPSCs into MNs and Purkinje cells using the 3D culture system we developed, as well as to detect some characteristic ARSACS pathological features such as abnormal accumulation of NFM along the neurites. Using collagen sponges populated with dermal fibroblasts as a 3D substrate, which is known to promote long-term survival and neurite elongation of MNs [ 39 ] and to reproduce pathological features observed in other brain diseases such as neurofibromatosis [ 46 ] and amyotrophic lateral sclerosis [ 47 , 48 ], it was possible to enable long term (>53 days) of Purkinje cells and to promote abundant elaborated dendritic ramifications. To better mimic ARSACS cellular microenvironment around MNs, iPSC-derived Schwann cells were also cocultured with iPSC-MNs within the 3D substrate.…”

Tissue Engineering for Gastrointestinal and Genitourinary Tracts

“…In this case, tissue engineering, especially the self-assembly method, can be helpful. For example, models have been developed to study skin pathologies such as hypertrophic scars [53], systemic sclerosis (scleroderma) [54], melanoma [55,56], psoriasis [57], epidermolysis bullosa [58], neurofibromatosis [59], or skin manifestations of amyotrophic lateral sclerosis (ALS) [60], but also genitourinary pathologies such as vaginal mucosa infection by the human immunodeficiency virus (HIV) [61], ketamine-induced cystitis [62], bladder cancer [63] or urinary tract infection [64].…”