Ksom aa

Manufactured by Merck Group

Sourced in United States

KSOM-AA is a culture medium used in assisted reproductive technologies. It is a modified version of the KSOM (Potassium Simplex Optimized Medium) formulation, designed to support the growth and development of embryos. KSOM-AA contains amino acids to supplement the culture environment.

Automatically generated - may contain errors

Lab products found in correlation

35 mm petri dishes, by BD (3 mentions) M2 medium, by Merck Group (3 mentions) Hyaluronidase, by Merck Group (3 mentions) Femtojet 4i microinjector, by Eppendorf (2 mentions) Embryomax, by Merck Group (2 mentions) C57bl 6j, by Jackson ImmunoResearch (2 mentions) Bovine serum albumin (bsa), by Merck Group (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Lsm 880, by Zeiss (2 mentions) High capacity reverse transcription kit, by Thermo Fisher Scientific (2 mentions) Acid tyrode, by Merck Group (2 mentions) Rneasy micro kit, by Qiagen (2 mentions) Chemidoc, by Bio-Rad (2 mentions) Micromanipulator, by Olympus (2 mentions) Femtojet microinjector, by Eppendorf (2 mentions) Cytochalasin b cb, by Merck Group (2 mentions) Heparin, by Merck Group (1 mentions) Fibroblast growth factor 2 (fgf2), by R&D Systems (1 mentions) Glass bottom dishes, by MatTek (1 mentions) Mr 070 d, by Merck Group (1 mentions)

Close spelling variants of this product

KSOM-AA medium EmbryoMax® KSOM + AA with D-glucose and phenol red

43 protocols using ksom aa

In Vitro Fertilization in Mice

Check if the same lab product or an alternative is used in the 5 most similar protocols

The female mice were superovulated by intraperitoneal injection of 5 IU pregnant mare serum gonadotropin (PMSG) and 5 IU of human chorionic gonadotropin (hCG) after 47 h. In the IVO group, the superovulated female mice were mated individually with male mice. The following morning, successful mating was confirmed by detecting the presence of a vaginal plug.
In the IVP group, IVF was performed as previously described [4 (link)]. Briefly, cumulus-enclosed oocyte complexes (COCs) were collected from the ampullae 14 h after the hCG injection, and the cumulus cells were removed by digesting the COCs with hyaluronidase for 3−5 min. The oocytes were first rinsed in human tubal fluid (HTF) medium (Sage, Bedminster, NJ, USA), and placed in 60-μL drops of HTF medium, covered with paraffin oil and equilibrated overnight at 37°C and 5% CO₂. Sperm was obtained from the cauda epididymis and capacitated for 1 h in HTF medium at 37°C and 5% CO₂. Sperm insemination was carried out 15 h after hCG injection. After 4 h in the incubator, the oocytes/embryos were washed several times in potassium simplex optimization medium containing amino acids (KSOM + AA; Millipore, Billerica, MA, USA) and transferred to 60-μL drops of KSOM + AA medium covered with paraffin oil. The embryos were cultured at 37°C with 5% CO₂.

Tan K., An L., Wang S.M., Wang X.D., Zhang Z.N., Miao K., Sui L.L., He S.Z., Nie J.Z., Wu Z.H, & Tian J.H. (2015). Actin Disorganization Plays a Vital Role in Impaired Embryonic Development of In Vitro-Produced Mouse Preimplantation Embryos. PLoS ONE, 10(6), e0130382.

+ Open protocol

+ Expand

Activation and Culture of Oocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

All treated oocytes were allowed to recover in a CO₂ incubator for 1 h before activation. The activation medium used was Ca²⁺-free human tubal fluid (HTF) [34 (link)] supplemented with 10 mmol/L SrCl₂ [35 (link)]. After being washed thrice in activation medium, oocytes were incubated first in activation medium for 2.5 h and then in regular HTF without SrCl₂ for 3.5 h at 37.5 °C in a humidified atmosphere with 5 % CO₂ in air. Both the activation medium and HTF for subsequent short culture of oocytes were supplemented with 2 μg/mL cytochalasin D. Six h after the onset of activation, oocytes were removed from the medium and cultured in KSOM-AA (simplex optimized medium contained K ions supplemented with amino acids) medium [36 (link)] (Millipore) for 4 d. Embryos at the two cell and blastocyst stages were examined and recorded at 24, and 96 h after start of culture in KSOM-AA medium, respectively.

Li W., Cheng K., Zhang Y., Meng Q., Zhu S, & Zhou G. (2015). No effect of exogenous melatonin on development of cryopreserved metaphase II oocytes in mouse. Journal of Animal Science and Biotechnology, 6(1), 42.

+ Open protocol

+ Expand

Generating Chimeric Mice via Embryo Aggregation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Chimeric mice have been created by either aggregation of cleaving embryos or injection of ESCs into cleaving host embryos. For aggregation, a pair of 4-8-cell stage ICR and NZB embryos were freed of zona pellucida, aggregated together in micro wells and cultured for 48hrs in AA-KSOM (Millipore) at 37°C and 5% CO2. Alternatively, 4–8-cell stage ICR host embryos were injected with NZB-Ubc-Td-tomato ESCs and 4–8-cell stage host NZB embryos were injected with B6-Ubc-GFP ESCs to generate chimeras. Injected embryos were cultured for 24 hrs in AA-KSOM medium (Millipore) at 37°C and 5% CO2 to blastocysts and then transferred to uteri of pseudo pregnant (E2.5 or E3.5) ICR females (recipients). Implantation sites were examined at gestation day 21.

Marti Gutierrez N., Mikhalchenko A., Ma H., Koski A., Li Y., Van Dyken C., Tippner-Hedges R., Yoon D., Liang D., Hayama T., Battaglia D., Kang E., Lee Y., Barnes A.P., Amato P, & Mitalipov S. (2022). Horizontal mtDNA transfer between cells is common during mouse development. iScience, 25(3), 103901.

+ Open protocol

+ Expand

Generating Mouse Models Using CRISPR

Check if the same lab product or an alternative is used in the 5 most similar protocols

Eight-week-old B6D2F1 female mice were superovulated with 6 international units of pregnant mare's serum gonadotropin (PMSG) for 48 h and then injected human chorionic gonadotropin (hCG), subsequently mated to homozygous Oct4-eGFP males for 12 h. Zygotes were harvested from oviducts of B6D2F1 females with plug 24 h post hCG injection using hyaluronidase (Sigma). For microinjection, the mixture of BE3 mRNA (100 ng/μl) and sgRNA (10, 20, 50 and 100 ng/μl) was diluted in RNAase-free water, centrifuged at 4°C, 13 400g for 10 min, and then injected into the cytoplasm of zygotes in a droplet of HCZB medium containing 5 μg/ml cytochalasin B (CB, Sigma) using a micromanipulator (Olympus) and a FemtoJet microinjector (Eppendorf). The injected zygotes were cultured for 16 or 24 h to two-cell embryos in AA-KSOM (Millipore) medium with different concentration of Ricolinostat or Nexturastat A, then cultured in normal AA-KSOM medium for 72 h to blastocyst stage at 37 °C under 5% CO₂ in air. Two-cell embryos were transferred into oviduct of pseudopregnant ICR females at 0.5 day post copulation for generating mouse models as previously described (23 (link)).

Zhao T., Li Q., Zhou C., Lv X., Liu H., Tu T., Tang N., Cheng Y., Liu X., Liu C., Zhao J., Song Z., Wang H., Li J, & Gu F. (2021). Small-molecule compounds boost genome-editing efficiency of cytosine base editor. Nucleic Acids Research, 49(15), 8974-8986.

+ Open protocol

+ Expand

Mouse Sperm Isolation and In Vitro Fertilization

Check if the same lab product or an alternative is used in the 5 most similar protocols

Spermatozoa collection and in vitro fertilization procedures were carried out as described⁵⁶. Briefly, sperm was isolated from the cauda epididymis of adult F1(C57BL/6×DBA) male mice and capacitated by pre-incubation for 1.5 h in pre-gassed HTF medium. In vitro matured oocytes were placed into HTF medium with capacitated sperm and incubated at 37 °C with 5% CO₂ atmosphere. After 4 h embryos were washed in 1:1 HTF:KSOMaa (Sigma-Alrich) for 20 min and further cultured in modified KSOMaa (supplemented with 3 mg/ml BSA, 5.4 mM glucose). For parthenogenetic activation, in vitro matured oocytes were washed three times with Ca²⁺-free CZB media and incubated for 20 min in Ca²⁺-free CZB media supplemented with 2.5 mM SrCl₂ at 37 °C with 5% CO₂ atmosphere. After SrCl₂ treatment, oocytes were washed three times in KSOMaa medium and cultured in modified KSOMaa medium at 37 °C with 5% CO₂ atmosphere. Oocytes were considered activated when each contained one (1PN) or two well-developed pronuclei (2PN). Parthenotes were fixed at 12 h post-activation.

Eleftheriou K., Peter A., Fedorenko I., Schmidt K., Wossidlo M, & Arand J. (2022). A transition phase in late mouse oogenesis impacts DNA methylation of the early embryo. Communications Biology, 5, 1047.

+ Open protocol

+ Expand

Live Imaging of Mouse Embryo Development

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Embryos were cultured in drops of KSOM-AA (Millipore) under mineral oil (Sigma) at 37°C in a 5% CO₂ atmosphere. For live imaging, embryos were cultured in glass-bottom dishes (MatTek) in an environmental chamber (Solent Scientific) as described previously (Piliszek et al., 2011 (link)). Embryos at post-cavitation stages were cultured without zona pellucidae. For FGF treatment experiments, FGF4 or FGF2 (R&D Systems) were added to culture medium at 500–2000 ng/ml, in the presence of 1 μg/ml heparin (Sigma). For ERK inhibition experiments 1μM of the ERK1/2 inhibitor PD0325901 (Stemgen) was added to the culture. Embryos were cultured from E2.5 or E3.5 various periods of time.

Schrode N., Saiz N., Di Talia S, & Hadjantonakis A.K. (2014). GATA6 Levels Modulate Primitive Endoderm Cell Fate Choice and Timing in the Mouse Blastocyst. Developmental cell, 29(4), 454-467.

+ Open protocol

+ Expand

In Vitro Fertilization of Mouse Oocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Twenty‐one days after surgery, the mice were injected intraperitoneally (ip) with 4 IU of eCG (Asuka Seiyaku) to stimulate preovulatory follicle development, followed by 5 IU of hCG (Asuka Seiyaku) 48 hours later to stimulate ovulation. In vitro fertilization was analyzed as described previously.²⁸ COCs that were collected from the oviduct 16 hours after hCG administration were placed in 50 μL of human tubal fluid (HTF) medium. Spermatozoa were collected from the cauda epididymis of each mouse genotype into 500 μL of HTF medium. After 60 minutes of incubation to induce sperm capacitation, the spermatozoa were introduced into the fertilization medium at a final concentration of 1000 spermatozoa/μL. Twelve hours after insemination, all gametes were further cultured for an additional day in the developing medium (KSOM + AA; Millipore) to check for development to the blastocyst stage.

Umehara T., Urabe N., Obata T., Yamaguchi T., Tanaka A, & Shimada M. (2020). Cutting the ovarian surface improves the responsiveness to exogenous hormonal treatment in aged mice. Reproductive Medicine and Biology, 19(4), 415-424.

+ Open protocol

+ Expand

Sperm Capacitation and Fertilization Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

IVF was done as described previously [27 (link)]. Cumulus-oocyte complexes (COCs) were collected from the oviduct of female mice at 16 hrs after an ovulatory hCG injection. Ten ovulated COCs collected from oviducts were placed in 50 μL of HTF medium for each IVF treatment.
Sperm were collected from the cauda epididymis to 1,000 μL of HTF medium or 0.3 μM TLR7/8 ligand–containing HTF medium. After 60 min of incubation to induce sperm capacitation and to react with TLR7/8 ligand, the upper layer (500 μL) was transferred to a new tube, and then was centrifuged at 37 °C for 5 min. The pellet of upper layer was suspended to HTF medium and was centrifuged at 37 °C for 5 min to remove the TLR7/8 ligand. On the other hands, the lower layer (300 μL) was also transferred to a new tube, and washed similar to the upper layer. After centrifuging, the pellet of sperm was suspended and transferred to fertilization medium at final number of 1,000 sperm per COC (S5 Fig). At 6 hrs after insemination, oocytes were examined for numbers of pro-nuclei to assess the fertilization and were further cultured in developing medium (KSOM+AA, Millipore, Billerica, MA, USA) to analyze blastocyst development. Some blastocysts were collected and were used to sexing of blastocysts using PCR.

Umehara T., Tsujita N, & Shimada M. (2019). Activation of Toll-like receptor 7/8 encoded by the X chromosome alters sperm motility and provides a novel simple technology for sexing sperm. PLoS Biology, 17(8), e3000398.

+ Open protocol

+ Expand

Zygote Microinjection of CRISPR Tools

Check if the same lab product or an alternative is used in the 5 most similar protocols

In preparation for zygote microinjection, C57BL/6J female mice at 4 weeks of age were superovulated by intraperitoneal injections of PMSG (5 IU, Prospec) and hCG hormone (5 IU, Sigma-Aldrich) with a 48-h interval between injections. For microinjection, a mixture containing left DdCBE (or DdCBE-NES)-encoding mRNA (300 ng/μl) and right DdCBE (or DdCBE-NES)-encoding mRNA (300 ng/μl) was diluted in DEPC-treated injection buffer (0.25 mM EDTA, 10 mM Tris, pH 7.4) and injected into the cytoplasm of zygotes using a Nikon ECLIPSE Ti micromanipulator and a FemtoJet 4i microinjector (Eppendorf). For co-injection of DdCBE and mitoTALEN, we added left and right mitoTALEN-encoding mRNAs (300 ng/μl each) to the injection buffer. After injection, embryos were cultured in microdrops of KSOM+AA (Millipore) at 37 °C for 4 days in a humidified atmosphere containing 5% CO₂. Two-cell stage embryos were implanted into the oviducts of 0.5-dpc pseudo-pregnant foster mothers.

Lee S., Lee H., Baek G., Namgung E., Park J.M., Kim S., Hong S, & Kim J.S. (2022). Enhanced mitochondrial DNA editing in mice using nuclear-exported TALE-linked deaminases and nucleases. Genome Biology, 23, 211.

+ Open protocol

+ Expand

In Vitro Fertilization and Embryo Culture

Check if the same lab product or an alternative is used in the 5 most similar protocols

In vivo or in vitro fertilized eggs (22–23 h after hCG), as determined by the presence of two pronuclei, were then cultured under optimized culture conditions (KSOM+AA, Millipore) and assessed for their developmental efficiency in vitro.

Chen S., Sun F.Z., Huang X., Wang X., Tang N., Zhu B, & Li B. (2015). Assisted reproduction causes placental maldevelopment and dysfunction linked to reduced fetal weight in mice. Scientific Reports, 5, 10596.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!