Questions? Feedback? powered by Olark live chat software
All Site

A straightforward and robust method of establishing 3-D Neurosphere Culture from Human iPSC-Derived Neural Progenitor Cells

Human induced pluripotent stem cell (iPSC)–derived neurospheres provide an advanced in vitro system for modeling the human brain. This system can be used to investigate areas of research focused on rare genetic diseases, disease pathology, immunological and host-pathogen interactions, and the toxic effects of environmental chemicals. Himanshi Desai et al. from ATCC use ATCC iPSC-derived neural progenitor cells (NPCs) to generate 3-D neurospheres in vitro. This report describes a straightforward and robust method of generating neurospheres and further supports their utilization for experimental applications such as dopaminergic differentiation and drug toxicity assays.

3-D cell culture systems, such as neurospheres, provide more robust platforms to study tissue-relevant physiology than traditional, 2-D models. In this study, they successfully generated 3-D neurospheres from normal- and Parkinson’s disease donor–derived NPCs. They first differentiated the NPCs into dopaminergic neurons using ATCC’s NPC Dopaminergic Differentiation Kit and subsequently cultured those neurons in ULA vessels until they aggregated into uniform circular spheres. Next, they assessed the potency of various chemotherapeutic drugs on neurospheres derived from normal or disease NPC lines. The method presented here is straightforward and reproducible because it does not require any synthetic scaffolds such as hydrogels or specialized equipment such as bioreactors. They believe that this method will provide the necessary flexibility to generate neurosphere cultures that fit the needs of specific research protocols.

Check which ATCC cells are used, protocols and experiment data in this report.

Ultra-low attachment (ULA) plates from Corning are used. Find plates and other plasticware required for both 3-D and 2-D cell culture.

celvivo banner 1


While you may be able to generate neurospheres using ULA plates, CelVivo has developed a clinostat incubator to create an environment which promotes the growth and maintenance of large 3D tissue mimetic structures, whether they are spheroids, organoids and other aggregates. The ClinoStar enables cell growth so they retain or recover their in vivo physiological attributes while obviating the need for scaffolds, gels or any type of additives which might perturb gene expression away from its natural baseline equilibrium.

rocio celvivo

Researchers in the Australian Institute for Bioengineering and Nanotechnology used the ClinoStar to culture brain organoids to better understand human ageing and its interplay with DNA damage, neurotoxicity and inflammation.



In collaboration with our partners (ATCCBio-techne, CelVivo, and Tantti), we are answering some of your biggest questions on 3-D cell culture. Find the top 20 3-D cell culture FAQS.

cell culture banner 3d