M2 medium

Manufactured by Eppendorf

Sourced in Germany

The M2 medium is a sterile, liquid culture medium designed for the growth and maintenance of a variety of cell types in cell culture applications. It provides the necessary nutrients and growth factors to support cell proliferation and viability.

Automatically generated - may contain errors

Lab products found in correlation

M2 medium, by Merck Group (2 mentions) Milrinone, by Merck Group (1 mentions) Milrinone, by Eppendorf (1 mentions) M16 medium, by Merck Group (1 mentions) Femtojet injector, by Eppendorf (1 mentions) B6cbaf1, by Janvier Labs (1 mentions)

2 protocols using m2 medium

Cytoplasmic Injection of siRNA in GV Oocytes

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Germinal Vesicle stage (GV) oocytes were collected from 5 to 8 weeks old B6CBAF1 (Janvier Labs, France) female mice at 39 h post-PMSG (Pregnant mare’s serum gonadotrophin, Intervet, France) injection. GV oocytes pick-up was performed in M2 medium (Sigma) supplemented with 2.5 μM Milrinone (Sigma) to maintain prophase I arrest. Cumulus cells were removed by repeated pipetting. Ten pl of siRNA solution were injected into the cytoplasm of denuded GV oocytes 1 hour after their retrieval, using a 1.2 µm inner diameter pipette (BioMedical Instruments, Germany) connected to a FemtoJet injector (Eppendorf, Germany) under a Nikon inverted microscope in the same M2 medium containing Milrinone at room temperature. Oocytes were then washed several times to remove Milrinone and matured in vitro in M16 medium (Sigma) during 24 or 48 h at 37°C, 5% CO₂ in air. siRNA against Ezrin (sc-35350), Radixin (sc-36367), Moesin (sc-35956), ERM (sc-37850) EWI-2 (sc-105564) and EWI-F (sc-142204) siRNA from Santa Cruz biotechnology were used at 1 μM final concentration. Control oocytes were injected following the same protocol with the same volume and concentration of control (CTRL) siRNA-A solution (sc-37007, Santa Cruz biotechnology). Figure 1 represents the experimental design of the study.

Cohen J., Wang L., Marques S., Ialy-Radio C., Barbaux S., Lefèvre B., Gourier C, & Ziyyat A. (2022). Oocyte ERM and EWI Proteins Are Involved in Mouse Fertilization. Frontiers in Cell and Developmental Biology, 10, 863729.

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Microinjection of Oocytes and Embryos

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Female mice were superovulated with pregnant mares serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG). MII oocytes were collected 14 h after hCG injection in M2 medium (Millipore) and freed of cumulus cells using hyaluronidase. For collection of 1-cell embryos, superovulated female mice were mated to BALB/c males and fertilized embryos were collected 18-20 h after hCG injection from oviduct. mRNA (50 – 250ng/μl) was injected into the cytoplasm of MII oocytes and fertilized embryos in M2 medium using Eppendorf micromanipulator. The injected MII oocytes were in vitro cultured in KSOM (Millipore) for 48 h before analysis. The injected embryos were cultured in KSOM at 37°C under 5% CO₂ in air until blastocyst stage. Subsequently, blastocysts were collected for analysis or transferred to BALB/c pseudopregnant females. Live pups were obtained by natural delivery.

Reddy P., Ocampo A., Suzuki K., Luo J., Bacman S.R., Williams S.L., Sugawara A., Okamura D., Tsunekawa Y., Wu J., Lam D., Xiong X., Montserrat N., Esteban C.R., Liu G.H., Sancho-Martinez I., Manau D., Civico S., Cardellach F., O’Callaghan M.D., Campistol J., Zhao H., Campistol J.M., Moraes C.T, & Belmonte J.C. (2015). Selective elimination of mitochondrial mutations in the germline by genome editing. Cell, 161(3), 459-469.

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