Table 1.

Characteristics of cNF in vitro models

		Cell Type
2D Versus. 3D	Species	Primary	Semi-Immortalized	iPSC or ESC derived	Source	Description	Comment	Reference
2D	Human	X			cNF, pNF	Methodology for expanding cNF SCs cultures by differential attachment		Wallace et al., 2000
							Used for testing in vivo tumorigenic properties of cNF-derived SCs	Muir et al., 2001
2D	Human	X			cNF	Methodology for expanding NFI+/− cNF SCs		Rosenbaum et al., 2000
2D	Human	X			cNF	Methodology for expanding NF1−/− cNF SCs		Serra et al., 2000
							Used for identifying somatic NF1 pathogenic variants	Maertens et al., 2006; Serra et al., 2001
							Used for genetic diagnosis of mosaic NF1 cases	Maertens et al., 2007
							Used for testing of hormones	Pennanen et al., 2018
							Drug screening	Serra laboratory Mazuelas et al.1
2D	Human	X			cNF	Cocultures of 70% of NF1−/− SC and 30% of NFI+/− FB	Drug screening	Serra laboratory Mazuelas et al.1
2D, 3D	pig	X			cNF	cNF-derived SCs and FBs from NF1R1947*/+ Ossabaw minipigs	Drug testing	Largaespada laboratory (unpublished data); Isakson et al., 2018
2D	Human		X		cNF	cNF-derived semi-immortalized SC lines	NFI+/−;NF1−/− immortalized SCs from cNF, similar to the pNF-immortalized SCs (Li et al., 2016)	Wallace laboratory; Wallace et al. (unpublished data)
2D	Human			X	iPSC derived	Differentiation of NF1+/+,NFI+/− and NF1−/− hiPSCs into SLCs	NF1+/+ and NFI+/− hiPSCs SLCs did not form tumors upon engraftment, whereas NF1−/− hiPSCs SLCs did	Mo et al., 2021
2D	Human			X	iPSC derived	NF1−/− edited/reprogrammed iPSCs differentiated into NCs and further to SCs	iPSC-derived NF1−/− differentiating SCs show a high proliferation and poor differentiation capacity in 2D	Carrió et al., 2019
2D	Human			X	ESC derived	SOX10:GFP reporter hESC line; NF1−/SMASh	In development Testing of SC behavior in response to different doses of NF1. Not suitable for drug testing.	Lee et al. (unpublished data)
2D	Human			X	ESC derived		In development Useful to study cNF pathogenesis and the effect of different levels of NF1 loss on SC development.	Lee et al. (unpublished data)
3D/organoid	Human	X			cNF	NFI+/−;NF1−/− cells seeded in Matrigel in miniring format	Setting up conditions Drug testing of 43 kinase inhibitors	Nguyen et al., 20222
3D	Human		X		cNF	cNF-derived semi-immortalized SC lines (Wallace et al., unpublished data)	MEKi tested had activity in 3D but not in 2D	Mattingly laboratory (unpublished data)
3D	Human			X	iPSC derived	NFI+/−;NF1−/− hiPSCs into SCs, alone or cocultured with cNF-derived primary fibroblasts	Drug testing	Serra laboratory Mazuelas et al.1
3D	Mouse	X			cNF	Spherogenic culture o Nf1−/− BC-derived stem-like cells	Drug screening (compared with MEKi) Mechanisms governing spontaneous LOH	Topilko laboratory Coulpier et al. (unpublished data)
3D/ organoid skin raft	Mouse	X			cNF	Nf1−/− skin neurosphere cells cocultured with Nfi+/− DRGs, sciatic nerves, and FBs on a layer of collagen type I	Interrogate interactions and spatial relations between different cell types; preclinical drug testing	Liao et al., 2016

Abbreviations: 2D, two-dimensional; 3D, three-dimensional; BC, boundary cap; cNF, cutaneous neurofibroma; DRG, dorsal root ganglion; ESC, embryonic stem cell; FB, fibroblast; hESC, human embryonic stem cell; hiPSC, human induced pluripotent stem cell; iPSC, induced pluripotent stem cell; LOH, loss of heterozygosity; MEKi, MAPK/extracellular signal—regulated kinase kinase inhibitor; NC, neural crest; NF1, neurofibromatosis type 1; pNF, plexiform neurofibroma; SLC, Schwannian lineage cell.