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Equafetal

Manufactured by Atlas Biologicals
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Sourced in United States, Japan

EquaFETAL is a laboratory instrument designed for the preparation and analysis of fetal bovine serum (FBS) samples. The core function of EquaFETAL is to provide a standardized and consistent method for the processing and evaluation of FBS, which is a widely used supplement in cell culture media.

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Lab products found in correlation

Streptomycin, by Thermo Fisher Scientific (8 mentions) Penicillin, by Thermo Fisher Scientific (6 mentions) Amphotericin b, by Thermo Fisher Scientific (6 mentions) Gentamicin, by Thermo Fisher Scientific (4 mentions) Ham s f12, by Thermo Fisher Scientific (4 mentions) Collagen 4 from human placenta, by Merck Group (4 mentions) Human endothelial sfm, by Thermo Fisher Scientific (4 mentions) Dulbecco s phosphate buffered saline, by Thermo Fisher Scientific (4 mentions) Medium 199, by Thermo Fisher Scientific (4 mentions) Liberase th, by Roche (4 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions) Tryple select, by Thermo Fisher Scientific (3 mentions) Ascorbic acid, by Avantor (3 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) Jimt 1, by Addexbio (2 mentions) Equafetal, by Corning (2 mentions) Glutamax, by Corning (2 mentions) Glutamax, by Thermo Fisher Scientific (2 mentions) Mda mb 231, by American Type Culture Collection (2 mentions)

22 protocols using equafetal

1

Plaque Assay for Detecting ZIKV in Mosquito Saliva

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In addition to RT-qPCR, salivary secretions from ZIKV-infected Ae. aegypti were assayed by plaque assay to confirm the presence of infectious virus particles. Briefly, saliva from individual mosquitoes were diluted in cell culture medium and plated on sub-confluent monolayers of Vero cells. Cells were incubated for 1 hr. at 37°C after which a semi-solid medium overlay (Dulbecco’s Modified Eagle Medium with 4% EquaFETAL, 2X Pen-Strep and 4 μg/mL Amphotericin B, mixed with an equal volume of 1.2% Tragacanth gum) (EquaFETAL, Atlas Biologicals, Fort Collins, USA) (Tragacanth, MP Biomedicals, Santa Ana, USA) was added. Cells were incubated for 4–5 days at 37oC in 5% CO2, stained with crystal violet, and plaques enumerated.
Armstrong P.M., Ehrlich H.Y., Magalhaes T., Miller M.R., Conway P.J., Bransfield A., Misencik M.J., Gloria-Soria A., Warren J.L., Andreadis T.G., Shepard J.J., Foy B.D., Pitzer V.E, & Brackney D.E. (2019). Successive Bloodmeals Enhance Virus Dissemination within Mosquitoes and Increase Transmission Potential. Nature microbiology, 5(2), 239-247.
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2

Plaque Assay for Detecting ZIKV in Mosquito Saliva

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In addition to RT-qPCR, salivary secretions from ZIKV-infected Ae. aegypti were assayed by plaque assay to confirm the presence of infectious virus particles. Briefly, saliva from individual mosquitoes were diluted in cell culture medium and plated on sub-confluent monolayers of Vero cells. Cells were incubated for 1 hr. at 37°C after which a semi-solid medium overlay (Dulbecco’s Modified Eagle Medium with 4% EquaFETAL, 2X Pen-Strep and 4 μg/mL Amphotericin B, mixed with an equal volume of 1.2% Tragacanth gum) (EquaFETAL, Atlas Biologicals, Fort Collins, USA) (Tragacanth, MP Biomedicals, Santa Ana, USA) was added. Cells were incubated for 4–5 days at 37oC in 5% CO2, stained with crystal violet, and plaques enumerated.
Armstrong P.M., Ehrlich H.Y., Magalhaes T., Miller M.R., Conway P.J., Bransfield A., Misencik M.J., Gloria-Soria A., Warren J.L., Andreadis T.G., Shepard J.J., Foy B.D., Pitzer V.E, & Brackney D.E. (2019). Successive Bloodmeals Enhance Virus Dissemination within Mosquitoes and Increase Transmission Potential. Nature microbiology, 5(2), 239-247.
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3

Fibroblast and LCL Cell Culture Conditions

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In this study, we used 24 type I SMA, 23 type II SMA, 11 type III SMA and 26 healthy control fibroblasts and LCLs. The lowest passage number stock vial was used for each cell line. All cell lines were maintained in a humidified 37°C incubator with 5% CO2. Fibroblasts were maintained in Dulbecco’s modified essential medium (DMEM; Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (EquaFETAL; Atlas Biologicals, Fort Collins, CO), 2 mM L-glutamine (Life Technologies) and 1% penicillin-streptomycin (Life Technologies). LCLs were maintained in RPMI-1640 medium (Life Technologies) supplemented with 15% EquaFETAL, 2 mM L-glutamine and 1% penicillin-streptomycin. Cell pellets were collected from each line within 3 passages.
Stabley D.L., Holbrook J., Harris A.W., Swoboda K.J., Crawford T.O., Sol-Church K, & Butchbach M.E. (2017). Establishing a reference dataset for the authentication of spinal muscular atrophy cell lines using STR profiling and digital PCR. Neuromuscular disorders : NMD, 27(5), 439-446.
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4

Comparative Analysis of Spinal Muscular Atrophy Fibroblasts

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Fibroblasts derived from type II SMA (GM03813, GM22592 and AIDHC-SP22) and non-SMA (GM03814, AIDHC-NMC1, AIDHC-SC1 and AIDHC-SC2) individuals were grown in DMEM containing 10% EquaFETAL (Atlas Biologicals, Fort Collins, CO), 2 mM L-glutamine (Life Technologies, Grand Island, NY) and 1% penicillin-streptomycin (Life Technologies). GM03813 [30 (link)], GM22592 and GM03814 [30 (link)] fibroblast lines were obtained from Coriell Cell Repositories (Camden, NJ) while the other fibroblast lines were generated at Nemours/Alfred I. duPont Hospital for Children. All type II SMA fibroblast lines used in this study contain 0 copies of SMN1 and 3 copies of SMN2 [31 (link)]. GM03814 fibroblasts [30 (link)] were derived from the carrier mother of GM03813 and contain 1 copy of SMN1 and 5 copies of SMN2 [31 (link)]. The other non-SMA fibroblast lines contain 2 copies of SMN1 and 2 copies of SMN2 [31 (link)]. The fibroblast lines were authenticated using short tandem repeat profiling and digital PCR as described previously [32 (link)].
The mouse motor neuron cell line NSC-34 [33 (link)] and the NSC-34-based reporter lines [18 (link);34 (link)] were maintained in DMEM, 5% EquaFETAL, 2 mM L-glutamine and 1% penicillin/streptomycin. In all instances, the cells were maintained in a humidified chamber at 37°C and 5% CO2.
Gentillon C., Connell A.J., Kirk R.W, & Butchbach M.E. (2017). The effects of C5-substituted 2,4-diaminoquinazolines on selected transcript expression in spinal muscular atrophy cells. PLoS ONE, 12(6), e0180657.
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5

Culturing Human Breast Cancer Cell Lines

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JIMT-1 (AddexBio), BT-474 (ATCC), and SKBR-3 (ATCC) were cultured in RPMI1640 (Corning) supplemented with 10% EquaFETAL® (Atlas Biologicals), GlutaMAX® (2 mM, Gibco), sodium pyruvate (1 mM, Corning), and penicillin-streptomycin (penicillin: 100 units mL–1; streptomycin: 100 µg mL–1, Gibco). KPL-4 (provided by Dr. Junichi Kurebayashi at Kawasaki Medical School)51 (link) and MDA-MB-231 (ATCC) were cultured in DMEM (Corning) supplemented with 10% EquaFETAL®, GlutaMAX®(2 mM), and penicillin-streptomycin (penicillin: 100 units mL–1; streptomycin: 100 µg mL–1). All cells were cultured at 37 °C under 5% CO2 and passaged before becoming fully confluent up to 10 passages. All cell lines were periodically tested for mycoplasma contamination. Cells were validated for the HER2 expression level in cell-based ELISA prior to use (see the following Cell-based ELISA section).
Anami Y., Yamazaki C.M., Xiong W., Gui X., Zhang N., An Z, & Tsuchikama K. (2018). Glutamic acid–valine–citrulline linkers ensure stability and efficacy of antibody–drug conjugates in mice. Nature Communications, 9, 2512.
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6

Cell Culture Conditions for Cancer Cell Lines

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JIMT-1 (AddexBio), JIMT-1(MDR1+) (generated in-house, see the protocol below), HCC1954 (ATCC), HCC1954-TDR (generated in-house, see the protocol below), SKBR-3 (ATCC), and THP-1 cells (ATCC) were cultured in RPMI1640 (Corning) supplemented with 10% EquaFETAL® (Atlas Biologicals), GlutaMAX® (2 mM, Gibco), sodium pyruvate (1 mM, Corning), and penicillin–streptomycin (penicillin: 100 units mL−1; streptomycin: 100 µg mL−1, Gibco). KPL-4 (provided by Dr. Junichi Kurebayashi at Kawasaki Medical School), MDA-MB-231 (ATCC), HepG2 (ATCC), and HEK293 (ATCC) were cultured in Dulbecco’s modified Eagle’s medium (Corning) supplemented with 10% EquaFETAL®, GlutaMAX® (2 mM), and penicillin–streptomycin (penicillin: 100 units mL−1; streptomycin: 100 µg mL−1). All cells were cultured at 37 °C under 5% CO2 and passaged before becoming fully confluent up to 20 passages. All cell lines were periodically tested for mycoplasma contamination. Cells were validated for the HER2 expression level in cell-based ELISA prior to use (see the “Cell-based ELISA assay” section).
Yamazaki C.M., Yamaguchi A., Anami Y., Xiong W., Otani Y., Lee J., Ueno N.T., Zhang N., An Z, & Tsuchikama K. (2021). Antibody-drug conjugates with dual payloads for combating breast tumor heterogeneity and drug resistance. Nature Communications, 12, 3528.
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7

Cell Line Maintenance and Genetic Modification

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All cell lines were maintained at 37°C, 5.0% CO2 in Gibco cell culture media: CHP212 (derived from male human neuroblastoma) in DMEM:F12 (1:1); 94T778 (derived from female human liposarcoma tissue) in DMEM:F12 (1:1); SJSA1 (derived from male human osteosarcoma) in RPMI-1640; A549 (derived from male human lung adenocarcinoma) in RPMI-1640; HEK293FT (derived from female human embryonic kidney tissue) in DMEM; MCF7 (derived from female human breast carcinoma metastasis) in DMEM; H4 (derived from male astrocytoma sample) in DMEM; HCT116 (derived from male human colorectal carcinoma tissue) in McCoy’s 5A. Cell lines identities were confirmed by STR analysis at the CU Anschutz Cell Technologies Shared Resource laboratory. Each medium was supplemented with 10% FBS (Peak Serum), except for 94T778 medium, which was supplemented with 10% EquaFetal (Atlas Biologicals), and 100 units/mL of penicillin, 100 μg/mL of streptomycin, and 250 ng/mL of amphotericin B (Gibco). Matching cultivation conditions were used for maintaining derived lineages: CHP212 stably transduced with 3xFLAG-FAM193A, HCT116 transiently transfected with Halo-FAM193A (full length and fragments 1–5) and MDM4-V5, and HEK293FT transiently transfected with Halo-FAM193A MDM4c3 (WT, deltaC, and C463A variants).
Szwarc M.M., Guarnieri A.L., Joshi M., Duc H.N., Laird M.C., Pandey A., Khanal S., Dohm E., Bui A.K., Sullivan K.D., Galbraith M.D., Andrysik Z, & Espinosa J.M. (2023). FAM193A is a positive regulator of p53 activity. Cell reports, 42(3), 112230.
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8

Myogenic differentiation of C2C12 cells

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Mouse C2C12 cells were provided by the RIKEN BioResource Research Center through the National BioResource Project of the MEXT/AMED (Ibaraki, Japan). The cells were cultured in the growth medium (GM) consisting of Dulbecco’s modified Eagle medium (DMEM) (Invitrogen, Life technologies Japan, Tokyo, Japan) supplemented with 10% fetal bovine serum substitute EquaFETAL (Atlas Biologicals, Fort Collins, CO, USA), at 37°C and 5% CO2 until the cells reached 60%–70% confluency. For each passage, the cells were trypsinized, diluted ten times, and seeded into new dishes for 72 h. Cells from the second passage were stored in a serum-free cell preservation medium Bambanker® (GC Lymphotec, Tokyo, Japan) at −80°C. For the experiments, the stock cells were thawed (passage 0) and passaging was repeated until a high passage number (passage 10 or passage 20) was achieved. Myogenic differentiation was induced by changing the GM to a differentiation medium (DM) consisting of 2% horse serum (InvitrogenTM) with DMEM. The DM was changed every other day. After 7 days, myotube formation was evaluated.
Shintani-Ishida K., Tsurumi R, & Ikegaya H. (2023). Decrease in the expression of muscle-specific miRNAs, miR-133a and miR-1, in myoblasts with replicative senescence. PLOS ONE, 18(1), e0280527.
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9

Isolation of Primary Myoblasts from Mouse Skeletal Muscle

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Primary myoblasts were isolated from the skeletal muscles of C57BL/6J male mice [22 (link)]. All methods were carried out in accordance with the Guidelines for Proper Conduct of Animal Experiments by the Science Council of Japan. The experimental protocol used in this study was reviewed and approved by the experimental animal committee of the Kyoto Prefectural University of Medicine (Permit No. M2020-1). The study was carried out in compliance with the ARRIVE guidelines. Skeletal muscle tissues of the hindlimbs were digested with collagenase II (Sigma-Aldrich Japan, Tokyo, Japan). Dissociated cells were suspended in Ham’s F-10 medium (InvitrogenTM) containing 20% EquaFETAL (Atlas Biologicals) and 2 ng/mL of human recombinant basic fibroblast growth factor (bFGF) (Nacalai Tesque, Kyoto, Japan). They were seeded onto noncoated dishes and incubated at 37°C in 5% CO2 overnight to remove the fibroblasts. The next day, the supernatant was collected and centrifuged. The pellets were resuspended in Ham’s F-10 medium, which contains 20% EqualFETAL and 2 ng/mL of bFGF, and cultured on collagen-coated dishes until 70%–80% confluent.
Shintani-Ishida K., Tsurumi R, & Ikegaya H. (2023). Decrease in the expression of muscle-specific miRNAs, miR-133a and miR-1, in myoblasts with replicative senescence. PLOS ONE, 18(1), e0280527.
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10

Isolation and Culture of Human Endothelial Cells

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Ham’s F12, Medium 199, Human Endothelial-SFM, fetal bovine serum (FBS), Dulbecco’s Phosphate-Buffered Saline (PBS), Insulin/Transferrin/Selenium (ITS), Collagenase Type I, TrypLETM Express (TE), TrypLETM Select (TS), gentamicin, amphotericin B, 5-ethynyl-29-deoxyuridine (EdU) incorporation Click-iT Alexa Fluor 488 cell proliferation assay kit, Fix and Perm (Medium A), penicillin and streptomycin were purchased from Life Technologies (Carlsbad, CA, USA). Dimethyl sulfoxide (DMSO), trypan blue (0.4%), alizarin red, paraformaldehyde (PFA), human serum (HS), Collagen IV from human placenta, ascorbic acid, and ascorbate-2-phosphate were purchased from Sigma (St. Louis, MO, USA). Recombinant basic fibroblast growth factor was bought from R&D Systems (bFGF, Minneapolis, MN, USA). Rho-associated, coiled-coil protein kinase inhibitor Y-27632 was brought from Miltenyi Biotec (Bergisch Gladbach, Germany). FNC coating mixture was obtained from United States Biologicals (Swampscott, MA, USA). Liberase TH was purchased from Roche (Mannhein, Germany). EquaFetal® was from Atlas Biologicals (Fort Collins, CO, USA). FREEZEstem cryo-preservation medium, LAMSCREENTM, human recombinant laminin-511 and 521 were bought from BioLamina (Sundbyberg, Sweden). Finally, Cryoscarless cryo-preservation medium was from Funakoshi (Tokyo, Japan).
Peh G.S., Ang H.P., Lwin C.N., Adnan K., George B.L., Seah X.Y., Lin S.J., Bhogal M., Liu Y.C., Tan D.T, & Mehta J.S. (2017). Regulatory Compliant Tissue-Engineered Human Corneal Endothelial Grafts Restore Corneal Function of Rabbits with Bullous Keratopathy. Scientific Reports, 7, 14149.
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