VitroGelĀ® Organoid Recovery Solution
(Improved Formulation over VitroGelĀ® Cell Recovery Solution)
Product Advantages
- Works both with cells cultured with VitroGelĀ® hydrogels or an animal-based ECM
- High recovery and cell viability
- Yields high-quality cells/organoids for downstream analysis or expansion
- 2-minute dissociation of animal-based ECM for intact organoids/cells
- <15 minute protocol for VitroGelĀ® hydrogels
- Enzyme-free formulation
- Stable for 15 months
- Easy-to-use with room temperature operation
- Longer shelf life than competitors. No cold pack shipping.
VitroGelĀ® Organoid Recovery Solution is a non-enzymatic cell harvesting solution for quick and efficient recovery of 3D cells or organoids cultured with either VitroGelĀ® hydrogels or an animal-based ECM.
For cells/organoids cultured in an animal-based ECM, achieve fast 2-minute ECM dissociation safely and efficiently.Ā
VitroGelĀ® Organoid Recovery Solution is room temperature stable and has a neutral pH. The solution can maintain high cell viability during the recovery process. Harvested cells can be subcultured in both 3D and 2D cultures. The VitroGelĀ® Organoid Recovery Solution can be used before or after the fixation and stained preparation of hydrogel specimens to ensure high-quality downstream data analysis.
VitroGelĀ® Organoid Recovery Solution compared to other recovery solutions
Specifications
Sizes | 100 mL, 500 mL |
Formulation | Enzyme-free. Free of phenol red. Sterile. |
Use | Harvest organoids/cells cultured from animal-based ECM or from VitroGel hydrogels. Use before or after sample fixation and stained preparation for imaging or downstream data analysis |
Processing Time | 10-15 min |
Downstream | Recovered cells can be sub-culture in both 3D and 2D culture |
pH | Neutral |
Storage | 2-8 Ā°C |
Stability | 15 months from date of manufacture |
Shipping | Ambient temperature |
Protocols / Resources
Video Protocols & Demonstrations
Organoid/Cell Recovery Protocol Workflows
From Animal-based ECM (e.g. Matrigel)
Less than 10 min protocol.Ā Fast 2-minute ECM dissociation.
Add recovery solution to the cells and transfer to a tube.
Rock tube for 2 minutes and centrifuge.
From VitroGelĀ® Hydrogels
Less than 15 min protocol.
Application Notes
Data and References
Figure 1.Ā Organoids recovered from Matrigel using VitroGelĀ® Organoid Recovery Solution with two methods.
Method 1: Re-suspend organoids in VitroGelĀ® Organoid Recovery Solution by pipetting to break organoids into small fragments for sub-culture/expansion.
Method 2: Rocking the tube with organoids and VitroGelĀ® Organoid Recovery Solution mixture without using a pipette to harvest the intact organoids.
In both, VitroGelĀ® Organoid Recovery Solution was kept in a 4Ā°C refrigerator to maintain a low temperature before use. The organoids/Matrigel and VitroGelĀ® Organoid Recovery Solution mixture were incubated at room temperature for 2 min before centrifuging. Day 0 images show the morphology of organoids right after harvesting with two different methods.
Figure 2.Ā Comparison of VitroGelĀ® Organoid Recovery Solution and VitroGel Cell Recovery Solution for organoids recovered from Matrigel.
Organoids (Passage 19) were cultured in Matrigel for three days before harvesting. 1 mL cold (4Ā°C) VitroGelĀ® Organoid Recovery Solution (MS04) or VitroGel Cell Recovery Solution (MS03) was added to a well of a 24-well plate to resuspend the organoid-Matrigel dome. The mixture was gently pipetted, transferred to a centrifuge tube, and incubated at room temperature for 2 min (MS04) and 10 min (MS03), respectively. After centrifuging to collect the organoids, the Matrigel in MS04 solution was completely dissolved. For the MS03 solution, there was a small amount of soft gel remaining on top of the organoids. The organoids harvested from both conditions were sub-cultured in Matrigel. The images above (Passage 20, Days 1&2) show the healthy organoid expansion from both cell recovery solutions. VitroGelĀ® Organoid Recovery Solution shows a higher recovery efficiency than the VitroGelĀ® Cell Recovery Solution in this study.
Figure 3.Ā Comparison of VitroGelĀ® Organoid Recovery Solution with other organoid recovery solutions.
The data above shows the comparison study of VitroGelĀ® Organoid Recovery Solution (MS04) with other organoid recovery solutions (Solution RLR from Company SC and Solution 3ODR from Company SA). All solutions were used directly from a 4Ā°C refrigerator. After mixing with organoid/Matrigel, the mixture with MS04 was incubated at room temperature for 2 min, while the mixture with Solution RLR or Solution 3ODR was incubated in an ice bucket for at least 15 min. After centrifuging, all Matrigel in the MS04 mixture was dissolved, but most Matrigel remained in both Solution RLR and Solution 3ODR conditions. Besides recovering intact organoids, the organoids from the MS04 solution can be easily resuspended in a cell culture medium and broken into small fragments for a subculture (MS04 images above). However, the organoid/Matrigel mixture was hard to break in Solution RLR or Solution 3ODR.Ā VitroGelĀ® Organoid Recovery Solution delivers a higher cell recovery rate for organoid expansion.
Figure 4.Ā Use VitroGelĀ® Organoid Recovery Solution for iPSC harvesting from 2D Matrigel coating plate
VitroGelĀ® Organoid Recovery Solution can be used to harvest iPSC cells from a 2D Matrigel coating plate.Ā The solution was warmed up to room temperature before use.Ā A) Morphology of cells detaching from the Matrigel coating plate. (3 min after adding VitroGelĀ® Organoid Recovery Solution), B) Image of the well plate after cell harvesting. (Shows all cells were removed from the Matrigel coating plate), C) Morphology of cells after re-seeding to a new Matrigel coating plate (Day 3).
References/Publications
- De Donato, M., Babini, G., Mozzetti, S., Buttarelli, M., Ciucci, A., Arduini, G., De Rosa, M. C., Scambia, G., & Gallo, D. (2020).Ā KLF7: a new candidate biomarker and therapeutic target for high-grade serous ovarian cancer.Ā Journal of Experimental & Clinical Cancer Research, 39(1). https://doi.org/10.1186/s13046-020-01775-9
- Lan, T., Guo, J., Bai, X., Huang, Z., Wei, Z., Du, G., Yan, G., Weng, L., & Yi, X. (2020).Ā RGD-modified injectable hydrogel maintains islet beta-cell survival and function.Ā Journal of Applied Biomaterials & Functional Materials,Ā 18, 228080002096347. https://doi.org/10.1177/2280800020963473
- Powell K. Adding depth to cell culture. Science, 356(6333), 96ā98. https://doi.org/10.1126/science.356.6333.96