NEW
Vial with white cap labeled 'RocketCell S3 Supplement (50X), AOF' — sterile, -20°C, 10 mL; REF RCS3, LOT 00000000; QR code; FOR RESEARCH USE ONLY
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Fluorescent image of a cell cluster with blue and magenta staining, shows a spherical cell aggregate on a black background.
Vial with white cap labeled 'RocketCell S3 Supplement (50X), AOF' — sterile, -20°C, 10 mL; REF RCS3, LOT 00000000; QR code; FOR RESEARCH USE ONLY
Abstract paint-splatter art: two bright yellow clusters with red, orange, green spokes radiating on a black background.
Fluorescent image of a cell cluster with blue and magenta staining, shows a spherical cell aggregate on a black background.

RocketCell™ S3 Supplement (50X), AOF

N2B27-Like Premixed Animal Origin-Free Supplement

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RocketCell™ S3 Supplement (50X), AOF

N2B27-Like Premixed Animal Origin-Free Supplement

Synergistic Dual-Supplement Performance
Combining the complementary strengths of both formulations delivers superior cell viability and performance compared to either supplement used alone.

3D and Co-culture Compatible
Formulated for stability and performance within VitroGel® hydrogel-based 3D culture systems, supporting complex co-culture and organoid model development.
Also works with animal-based ECMs.

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Consistent Lot-to-Lot Quality
Selectively supports post mitotic neurons in mixed primary cultures while suppressing overgrowth of non-neuronal cell populations.

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Ready-to-use Convenience
Eliminates the manual mixing step, reducing pipetting variability and preparation time in high-throughput or time-sensitive workflows.

Broad Application Range
Supports neural stem cell expansion, iPSC-derived neuron maturation, cancer stem cell maintenance, and organoid and tumoroid culture across multiple tissue types including intestinal, colonic, pancreatic, lung, retinal, and liver models.

RocketCell™ S3 Supplement, AOF (N2B27) is animal origin-free, ready-to-use, 1:1 premix of the RocketCell™ S2 Supplement (N2-like) and S1 Supplement (B27-like) formulations — a single-component solution for culture paradigms that require both supplements simultaneously.

The combination of N2 and B27 is a cornerstone of modern serum-free culture, appearing in media formulations across an exceptionally broad range of applications including neural, epithelial, and cancer models, as well as organoid and tumoroid systems across multiple tissue types. RocketCell™ S3 Supplement, AOF delivers this established dual-supplement framework in a fully animal origin-free and defined format.

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Challenges We Solve

Common Challenges 

Lack of a reliable animal origin-free formulation to support GMP process.

Multi-supplement media preparation increases preparation time and handling steps​.

Variability in media preparation when multiple supplements are mixed manually​.

Limited availability of ready-to-use N2/B27-like supplements for diverse organoid models​.

How RocketCell™ S3 Supplement, AOF Solves The Issues​

Animal origin-free formulation for GMP transition
N2/B27-like AOF formulation that simply the transition to
scalable and controlled GMP workflows. ​

Simplified Media Preparation
Combines N2- and B27-like components in one bottle, reducing media prep time, pipetting steps, and contamination risk.

Three stacked rounded rectangles in a row with a checkmark circle above, symbolizing completion or approval of all three items.

Consistent Supplementation
Delivers a standardized N2B27-like formulation across
experiments and workflows.​

Robust Formulation for Organoid Applications
Supports neural stem cell expansion, iPSC-derived neuron maturation, cancer stem cell maintenance, and organoid/tumoroid culture across multiple tissue types.

Data and References

iPSC-derived Intestinal Organoids Cultured Using RocketCell™ S3 (N2B27) Supplement, AOF

DAPI staining highlights nuclei in cyan within a cell cluster (panel A).","Phalloidin labeling shows actin-rich structures in yellow (panel B).","Villin staining reveals magenta microvilli-rich cytoplasm (panel C).","Merged multicolor image combines DAPI, Phalloidin, and Villin signals (panel D).

Use of VitroGel® hydrogels & RocketCell™ S3 Supplement, AOF to generate iPSC-derived intestinal organoids with apical-out polarity.

Representative image showing iPSC derived intestinal organoids cultured in VitroGel® ORGANOID. iPSC cells were cultured in VitroGel® STEM to generation of spheroids cells. These spheroids were then differentiated to intestinal organoids using organoid culture media formulated using RocketCell™S3 Supplement, AOF and CytoGrow™ growth factors. (A) Blue staining (DAPI) represents cell nuclei. (B) Yellow fluorescence: Phalloidin, (C) Magenta: Villin protein, & (D) Merged image.​

Generation of iPSC-derived Liver Organoid in RocketCell™ S3 (N2B27) Supplement, AOF and Synthetic VitroGel® ORGANOID Hydrogel

Panel of three rows and three columns showing fluorescence with DAPI (red) and specific markers (green, blue, magenta, orange) and merged images. Row 1: DAPI red nuclei (A1), HNFa4 green (B1), merged (C1). Row 2: DAPI red (A2), PCK2 blue (B2), merged (C2). Row 3: DAPI red nuclei (A3), SOX9 green (B3), Phalloidin-magenta/merged (C3).

Generation and characterization of iPSC-derived liver organoids using RocketCell™ S3 (N2B27) Supplement and synthetic, chemically defined VitroGel® ORGANOID Hydrogel.

Representative bright-field images demonstrate the formation and progressive development of iPSC-derived liver organoids in 3D culture. Immunofluorescence analysis confirms liver-specific characteristics, including HNF4α expression (1), PCK2 expression and F-actin/Phalloidin staining (2), and SOX9 expression (3). DAPI staining indicates cell nuclei, with merged images showing the corresponding marker expression within the organoid structures.

Mouse Intestinal Organoid Development using
RocketCell™ S3 (N2B27) Supplement, AOF in Animal-based ECM​

Panel A: Day 0 culture in RocketCell S3 Supplement (AOF) in a round dish with dense cell material scattered across the surface.
Line chart of mouse intestinal organoid growth over 5 days. Blue line with markers shows increasing organoid area (normalized) with error bars; gray line shows a slower increase for Commercial Intestinal Organoid Media. Legend indicates blue = Media with RocketCell S3 Supplement, AOF; gray = Commercial Intestinal Organoid Media. X-axis: Time (Days) from 0 to ~4.5; Y-axis: Organoid Area (normalized).

Mouse intestinal organoid growth in animal-based ECM using RocketCell™ S3 Supplement, AOF.

Representative bright-field images show organoid development from Day 0 to Day 5 in RocketCell™-supplemented medium compared with commercial intestinal organoid medium. Quantitative analysis demonstrates enhanced organoid growth with RocketCell™ medium, with greater expansion observed from approximately Day 2 onward.​

Differentiated Neurospheres (Brain Organoids) developed in VitroGel® NEURON, RocketCell™ NSC Growth Medium, AOF and RocketCell™ S3 (N2B27) Supplement, AOF​

Multichannel microscopy grid: blue DAPI nuclei, green MAP2 neurites, red NF-L, magenta Synaptophysin, plus depth-shaded and phase images.

Neurosphere differentiation and culture in VitroGel® NEURON, RocketCell™ NSC Growth Medium, AOF, and  RocketCell™ S3 Supplement , AOF

Immunofluorescence staining of differentiated Neuron-Spheres (Brain Organoids) developed in 3D using VitroGel® NEURON and RocketCell™ NSC Growth Medium, AOF in combination with RocketCell™ S3 Supplement, AOF (N2B27 Alternative). The images were obtained with Confocal Microscopy – Maximum Projection and Incucyte S3 Standard 20x Tracking​.

Neurospheres (Brain Organoids) Differentiation in VitroGel® NEURON, RocketCell™ NSC Growth Medium, AOF and ​RocketCell™ S3 (N2B27) Supplement, AOF​

Quad-panel fluorescence microscopy showing marker maps of neural cells. Top-left: 3D image with Nestin (purple/magenta) and DAPI (blue) staining, large cell cluster, 50 μm scale bar. Top-right: 2D image with Nestin (green) and Sox2 (magenta), dense progenitor network, 100 μm scale bar. Bottom-left: 2D image of Sox2 staining in red. Bottom-right: 2D image of Pax6 staining in magenta.

Growth of NSCs in VitroGel® STEM using RocketCell™ NSC Growth Medium, AOF

​iPSC-derived NSCs were cultured in VitroGel® STEM and the RocketCell™ NSC Growth Medium, AOF supplemented with the RocketCell™ S3 Supplement, AOF (N2B27 Alternative) for 14 days. Immunofluorescence staining depicts expression of NSC-associated markers. ​

Size

10 mL

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