Continuous Single Culture
Single-step culture medium that reduces stress by eliminating unnecessary steps.
Further Information
Continuous Single Culture product (CSCM) is a novel solution designed for fertilization and embryo culture through day 5/6 of embryo development. CSCM is optimized to be used in an uninterrupted culture system without dish change or medium renewal.
CSCM is bicarbonate-based and is designed for use in a CO2 incubator.
Requires protein supplementation.
Available in 60mL configuration.
Storage: Store at 2-8°C
Shelf Life: 90 days from date of manufacture
| Size | SKU |
|---|---|
| 60mL | 90164-60mL |
Product Sheets
Protocols
White Papers & Research Articles
Scientific Posters
Presentations
Our complete media contain the concentration of protein indicated below; it is possible to add more protein if needed, as long as you consider that the following has already been added.
- Multipurpose Handling Media-Complete (MHM-C) with Gentamicin is pre-supplemented with HSA for a final protein concentration of 5 mg/mL.
- Continuous Single Culture-NX Complete (CSCM-NXC) is pre-supplemented with HSA for a final protein concentration of 5 mg/mL.
- Continuous Single Culture Complete (CSCM-C) with HSA is pre-supplemented with HSA for a final protein concentration of 5 mg/mL.
CSCM-NX and CSCM-NXC are clinically-proven, low lactate; single-step culture media that help improve blastocyst development. CSCM-NX and CSCM-NXC are designed for fertilization and embryo culture through day 5/6 of embryo development. These media are optimized to be used in an uninterrupted culture system, without dish change or medium renewal.
No, all Irvine Scientific fertilization and embryo culture media contain gentamicin as an antibiotic. All the Continuous Single Culture media (CSCM, CSCM-C, CSCM-NX, and CSCM-NXC) are formulated with 10 μg/mL gentamicin.
No, our expiration dates are based on our stability studies and we cannot guarantee product performance past the date of expiration. We recommend the customer to count back from the final day of culture and not use the product if the expiration date falls before the culture period is complete.
Yes, you can use your lab's protocol with all the Continuous Single Culture media (CSCM, CSCM-C, CSCM-NX, and CSCM-NXC); however, our data supports improved implantation and blastocyst utilization rates with an uninterrupted culture system which reduces embryo stress by eliminating the need for embryos to be exposed to varying media.
Protocols
- Uninterrupted Culture Media (CSCM-NX and CSCM) Recommended Protocol 4/5-Well Dish
- Uninterrupted Culture Media (CSCM-NX and CSCM) Recommended Protocol Microdrop
- Uninterrupted Culture Media Complete (CSCM-NXC and CSCM-C) Recommended Protocol 4/5-Well Dish
- Uninterrupted Culture Media Complete (CSCM-NXC and CSCM-C) Recommended Protocol Microdrop
Brochures
- From Gametes To Blastocysts
- The Best Possible Start - Continuous Single Culture-NX helps embryos minimize metabolic stress
When using Nexpring Health HSA, a 100 mg/mL protein solution, we recommend 5% v/v for a final concentration of 5 mg/mL. For example, to prepare 10 mL of medium, add 0.5 mL of HSA solution to 9.5 mL of the medium. When using Nexpring Health SSS, a 60 mg/mL protein solution, we recommend 10% v/v for a final concentration of 6 mg/mL. For example, to prepare 10 mL of medium, add 1.0 mL SSS to 9.0 mL of medium.
No, currently we do not use the MEGA assay as a final release test of Irvine Scientific culture media. The MEGA assay is currently only used for critical raw materials, such as Human Serum Albumin (HSA).
The working pH range for all the Continuous Single Culture media (CSCM, CSCM-C, CSCM-NX, and CSCM-NXC) is 7.25-7.40 with a targeted pH of 7.26-7.35.
Phenol red does not contribute to the performance of the culture medium and serves only as a visual indicator of pH (1). We recommend that CSCM, CSCM-C, CSCM-NX, and CSCM-NXC be equilibrated in a CO2 incubator with verified pH measurements prior to use. We will continue to offer CSCM and CSCM-C with phenol red.
References
- Berthois, Y., Katzenellenbogen, J. A., & Katzenellenbogen, B. S. (1986). Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture. Proceedings of the National Academy of Sciences,83(8), 2496-2500. doi:10.1073/pnas.83.8.2496
- Guerif, F., Mckeegan, P., Leese, H. J., & Sturmey, R. G. (2013). A Simple Approach for COnsumption and RElease (CORE) Analysis of Metabolic Activity in Single Mammalian Embryos. PLoS ONE,8(8). doi:10.1371/journal.pone.0067834
- Zander-Fox, D. & Lane, M. The future of human embryo culture media – or have we reached the ceiling? in The Human Embryo (ed. Yamada, S. & Takakuwa, T.) 73-98 (Intech, 2012).
Like CSCM and CSCM-C, CSCM-NX and CSCM-NXC are formulated using a bicarbonate buffering system that maintains a neutral system by balancing the presence of both a weak acid and its conjugate base. Moreover, phenol red is not a sensitive indicator of pH and does not participate in controlling the pH of the culture medium. Accordingly, pH is maintained in the recommended working range under the proper CO2 concentration.
No, all the Continuous Single Culture media (CSCM-NX, CSCM-NXC, CSCM, and CSCM-C) use the same recommended uninterrupted culture media protocol. Depending on a laboratory’s preference, a microdrop or well-based protocol can be used.
Protocols
- Uninterrupted Culture Media (CSCM-NX and CSCM) Recommended Protocol 4/5-Well Dish
- Uninterrupted Culture Media (CSCM-NX and CSCM) Recommended Protocol Microdrop
- Uninterrupted Culture Media Complete (CSCM-NXC and CSCM-C) Recommended Protocol 4/5-Well Dish
- Uninterrupted Culture Media Complete (CSCM-NXC and CSCM-C) Recommended Protocol Microdrop
The shelf life of CSCM-NX and CSCM-NXC was reduced to four (4) weeks based on the maximum length of time product is used in a laboratory after opening under aseptic conditions.
The lactate concentration of CSCM-NX and CSCM-NXC is 1 mM.
The reduction of lactate concentration in CSCM-NX and CSCM-NXC is a result of both MEA and BEA assays that support optimal blastocyst development in the presence of lower lactate concentrations. Lactate is a natural by-product of glucose metabolism produced by the embryo. Lowering the lactate concentration of CSCM-NX effectively reduces metabolic stress on the embryo and promotes efficient use of energy sources by the embryo (2,3).
References
- Berthois, Y., Katzenellenbogen, J. A., & Katzenellenbogen, B. S. (1986). Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture. Proceedings of the National Academy of Sciences,83(8), 2496-2500. doi:10.1073/pnas.83.8.2496
- Guerif, F., Mckeegan, P., Leese, H. J., & Sturmey, R. G. (2013). A Simple Approach for COnsumption and RElease (CORE) Analysis of Metabolic Activity in Single Mammalian Embryos. PLoS ONE,8(8). doi:10.1371/journal.pone.0067834
- Zander-Fox, D. & Lane, M. The future of human embryo culture media – or have we reached the ceiling? in The Human Embryo (ed. Yamada, S. & Takakuwa, T.) 73-98 (Intech, 2012).
Protein supplements contain human source material which can develop particulate matter during storage. When using protein-supplemented media, it is not uncommon to have particles form over time as a result of multiple warming and cooling cycles during their shelf life. This type of particulate matter does not adversely affect the performance of the media or embryo development.
These are the recommended target pH ranges for media with protein during use in a CO2 incubator. Typically the addition of protein will lower the pH by 0.02–0.05 units, but it should be within the ranges indicated as follows at 37°C and in a CO2 incubator.
- Human Tubal Fluid (HTF): 7.20 - 7.30
- Continuous Single Culture Medium (CSCM): 7.26 – 7.35
- Continuous Single Culture-NX (CSCM-NX): 7.26 – 7.35
Protocols
CSCM and CSCM-NX contain both essential and non-essential amino acids.
No, as indicated in the product insert the storage recommendation is between 2°C to 8°C. The product is stable and ready to use until the date shown on the bottle when stored unopened within that temperature range. The media should not be exposed to extreme temperatures at any time and, if frozen, we recommend discontinuing the use of the product. Please refer to individual product inserts for recommended storage and stability information.
Yes, culture media is offered with protein (complete media) or without protein.
| Catalog # | Culture Medium | Protein Source | Protein Concentration |
|---|---|---|---|
| 90165 | Continuous Single Culture Complete | HSA | 5 mg/mL |
| 90166 | Multipurpose Handling Media-Complete (MHM-C) with Gentamicin | HSA | 5 mg/mL |
| 90168 | Continuous Single Culture-NX Complete | HSA | 5 mg/mL |
The addition of EDTA has been shown to enhance the rate of early cleavage in embryos in culture and also serves as an antioxidant to stabilize the media.
Yes it is. Azide is commonly used as a preservative for diagnostic grade albumins.
Yes. HSA consists of human serum albumin from therapeutic-grade source material (100mg/mL) in a saline solution.
We offer both a light and heavy mineral oil intended for use as an overlay during embryo culture to prevent evaporation, changes in osmolality, and pH shifts.
