SpheroTribe provides a simple toolkit to generate consistent and robust 3D cell structures (spheroids, organoids) without restriction of cell type. Simply dilute the SpheroTribe solution into your culture medium of choice, watch your cells turn into uniformly sized 3D spheroids and collect them for your downstream assays.
Once diluted in your culture medium of choice, our concentrated polymer-based solution increases the medium viscosity favouring cell-cell contacts. SpheroTribe offers a simple method to generate homogeneous 3D cell structures with increased control over their size and shape, which can be easily handled and washed for downstream experiments.
SpheroTribe is particularly useful to boost aggregation when working with challenging cells, minimize variability between samples and improve the consistency of your migration/invasion assays, immunostaining, drug screening or in vivo implantation experiments.
Sizes:
- 10x 96-well plates (flat bottom), 2x tip boxes, 1 methylcellulose bottle (TDA-SPK-KIT)
- 1x 96-well plates (flat bottom), 20 tips, 2.5 mL of methylcellulose (TDA-SPK-MINIKIT)
- 1 methylcellulose bottle (TDA-SPK-2-25)
Catalogue Numbers:
- TDA-SPK-KIT (10x 96-well plates (flat bottom), 2x tip boxes, 1 methylcellulose bottle)
- TDA-SPK-MINIKIT (1x 96-well plates (flat bottom), 20 tips, 2.5 mL of methylcellulose)
- TDA-SPK-2-25 (1 methylcellulose bottle)
Kit description
In addition to the SpheroTribe solution, the full kit also includes U-bottom plates and wide-opening tips so you have everything you need to get started with your 3D cell culture experiments.
25mL kit (960 spheroids):
- 25mL of 5X methylcellulose solution
- 10x U-bottom 96-well plates
- 2x racks of 96 pipette tips (200µL) with a large opening
2,5mL kit (96 spheroids):
- 2.5mL of 5X methylcellulose solution
- 1x U-bottom 96-well plate
- 20 pipette tips (200µL) with a large opening
Storage recommendation for the methylcellulose solution: 4°C for (at least) 6 months.
Cell aggregation booster
SpheroTribe provides a gel-like scaffold that favors cell-cell contacts by increasing medium viscosity. It was shown to generate compact spheroids mimicking solid tumors, improve spheroid formation with some of the most challenging cells and to speed up stem cell-derived organoid formation.
Increased homogeneity
By maximizing cell aggregation, SpheroTribe promotes the formation of unique & uniformly sized spheroids allowing for consistent assays (growth, invasion, immune infiltration, in vivo injection, etc).
Easy to use
No need to work on ice, have access to sophisticated equipment of expertise. With SpheroTribe, you have everything on hand to easily grow & handle your spheroids.
Universal
The SpheroTribe solution is composed of methylcellulose, a biologically inert compound dissolved in basal culture medium without any proteins, lipids or growth factors. You can dilute it in any culture medium of your choice, and add additional compounds as desired (i.e. serum, antibiotics, differentiation factors, etc).
Applications:
So far, SpheroTribe has been successfully used for spheroid/organoid formation with the following cell types:
Patient-derived stem-like glioblastoma cells (GB P3, BL13, T033 & T042), human glioblastoma cell lines (U87 & T98G), HeLa, human vaginal mucosal melanoma (HMV-II), human primary colorectal cancer cells, human breast cancer cells (MDA-MB 231), human induced pluripotent stem cells, monkey kidney fibroblast-like cell line (COS-7), primary neurons from rat embryos (E18) & murine melanoma cells (B16F10).
Experimental assays:
Once spheroids have grown to your desired size, you can use them for any kind of assay according to your regular workflow. The SpheroTribe solution can be readily washed off, leaving a spheroid available for other tests at any stage of your protocol.
Example of in situ assays you can perform directly on the U-bottom plate supplied:
- Live imaging
- Growth/proliferation studies
- Toxicity studies
Examples of downstream assays that might require transferring spheroids to other vessels:
- Invasion and migration assays
- Immunostaining
- Biochemical assays
- Immune infiltration assays
- In vivo implantation
For more guidance on downstream assays, check out our FAQ section containing useful tips & example protocols.
Additional resources:
Results:
Uniform patient-derived glioblastoma spheroids generated using the SpheroTribe kit and visible by the naked eye after 4 days of culture
SpheroTribe improves the formation of unique & circular human glioblastoma spheroids
Human glioblastoma U87 cells were cultured in DMEM with or without SpheroTribe in U-bottom plates and imaged after 2 days (4X magnification) and 6 days (10X magnification)
Glioblastoma spheroids were included into a collagen matrix after being cultured in medium added with SpheroTribe solution for 4 days. Images were taken immediately after (left) or 24 hours after (right) inclusion in the collagen matrix.
Immune infiltration of B16F10 spheroids after immune checkpoint blockade
A. 10,000 B16F10 cells were grown for 6 days as spheroids using SpheroTribe. B. 100,000 PBMC from murine spleen were activated with IL-15 (40 ng/mL) [1], incubated with anti-PD1 (10 µg/mL) for 1h and added on B16F10 spheroids for 3 days. Graph shows flow cytometry quantification of differential lymphocyte infiltration after spheroid dissociation according to treatment. N=4. Mann-Whitney U Test, p-value<0.05. [1] https://doi.org/10.3389/fonc.2022.898732.
Image credits: Guillaume Mestrallet, PhD - Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, US - 2023.
SpheroTribe improves cardiac organoid formation by boosting hiPSC aggregation
Human iPSCs were seeded in ULA plates in presence (left) or absence (right) of SpheroTribe solution (T=0) and cultured for 3 days following a self-assembling human heart organoid differentiation protocol [1].
Image credits: Aitor Aguirre, Ph. D. - Michigan State University, US
[1] Lewis-Israeli, Y.R., Wasserman, A.H., Gabalski, M.A. et al. Self-assembling human heart organoids for the modeling of cardiac development and congenital heart disease. Nat Commun 12, 5142 (2021). https://doi.org/10.1038/s41467-021-25329-5
SpheroTribe improves spheroid formation in agarose micro-wells
U-87 glioblastoma cells were seeded at 1,000 cells per well in round-bottom wells molded in agarose using a Stampwell U-shape (Idylle) in full culture medium without (right) or added with SpheroTribe (left). After 3 days, pictures were taken and number of cell aggregates per well were quantified from 64 independent wells and associated standard deviation (SD) values were calculated.
SpheroTribe promotes and maintains cell aggregation in COS-7 monkey kidney fibroblast-like cells
COS-7 cells were cultured in non-tissue culture treated U-bottom plates for 4 days in presence (up)
or absence (bottom) of the SpheroTribe concentrate. Scale bar = 500 µm
SpheroTribe promotes and maintains cell aggregation in HeLa cells
HeLa cells were cultured in non-tissue culture treated U-bottom plates for 4 days in presence (up) or absence (bottom) of the SpheroTribe concentrate. Scale bar = 500 µm
How to use SpheroTribe in pictures
