Advanced minimum essential medium

Manufactured by Thermo Fisher Scientific

Sourced in United States

Advanced Minimum Essential Medium is a cell culture medium designed to support the growth and maintenance of a variety of cell types. It provides a balanced formulation of essential nutrients, vitamins, amino acids, and other components required for optimal cell culture performance.

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Lab products found in correlation

Glutamax, by Thermo Fisher Scientific (6 mentions) Penicillin, by Corning (2 mentions) Streptomycin, by Corning (2 mentions) Penn strep, by Corning (2 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions) Collagenase type 1, by Worthington (2 mentions) 70 μm cell strainer, by BD (2 mentions) Fetal bovine serum (fbs), by R&D Systems (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Collagenase 4, by STEMCELL (1 mentions) Pltgold human platelet lysate, by Merck Group (1 mentions) Gibco cell culture freezing medium, by Thermo Fisher Scientific (1 mentions) Trypsin like enzyme, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Collagenase type 1, by Thermo Fisher Scientific (1 mentions) Celltiter 96 non radioactive cell proliferation assay, by Promega (1 mentions) 100 μm cell strainer, by BD (1 mentions)

5 protocols using advanced minimum essential medium

Isolation and Propagation of Human MSCs

Cited 4 times

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Following informed consent and IRB approval at the authors’ institution, human MSCs were isolated from lipo-aspirates obtained from three representative donors (cell lines: 211, 258 and 283) using previously described methods [48 (link)–50 (link)]. The donor-derived MSCs selected in this study were propagated with platelet lysate containing zoonotic free media and have been previously shown to express standard MSC markers and to undergo multi-lineage differentiation [48 (link), 49 (link), 51 (link), 52 (link)]. In brief, lipo-aspirates were digested with Type I collagenase (Worthington Biochemicals, Lakewood, NJ) for 1.5 hours at 37°C, centrifuged at 400g for 5 min, rinsed with phosphate buffered saline (Life Technologies, Grand Island, NY), and strained using 70μm cell strainers (BD Biosciences, San Jose, CA). Following this, the aspirate was treated with 154 mM NH₄Cl, 10 mM KHCO₃, and 0.1 mM EDTA for erythrocyte lysis. The remaining MSCs were expanded in advanced minimum essential medium (Life Technologies, Grand Island, NY), supplemented with 5% (vol/vol) human platelet lysate (PLTMax; MillCreekLifeSciences, Rochester, MN), 2mM Glutamax (Life Technologies, Grand Island, NY), 2 U/mL heparin, and 1% Penn-Strep (100 U/mL penicillin, 100 μg/mL streptomycin; Cellgro, Corning, NY). All MSCs used in experimentation were of passage 7.

Nie H., Kubrova E., Wu T., Denbeigh J.M., Hunt C., Dietz A.B., Smith J., Qu W, & van Wijnen A.J. (2019). Effect of lidocaine on viability and gene expression of human adipose derived mesenchymal stem cells: an in vitro study. PM & R : the journal of injury, function, and rehabilitation, 11(11), 1218-1227.

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AMSC Isolation from Lipoaspirates

Cited 3 times

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Following informed consent and IRB approval at the authors′ institution, human AMSCs were isolated from lipo-aspirates obtained from a representative donor using previously described methods^{13 (link), 14 (link),49 (link)}. The donor-derived AMSCs selected in this study have previously been shown to express standard MSC markers and are capable of multi-lineage differentiation^{13 (link)–15 (link)}. In brief, donor lipo-tissue was aspirated and then digested using Type I collagenase (Worthington Biochemicals, Lakewood, NJ) for 1.5 hours at 37°C, centrifuged at 400g for 5 min, rinsed with phosphate buffered saline (Life Technologies, Grand Island, NY), and strained using 70μm cell strainers (BD Biosciences, San Jose, CA). Following this, the aspirate was treated with 154 mM NH₄Cl, 10 mM KHCO₃, and 0.1 mM EDTA for erythrocyte lysis. The remaining AMSCs were expanded in advanced minimum essential medium (Life Technologies, Grand Island, NY), supplemented with 5% (vol/vol) human platelet lysate (PLTMax; MillCreekLifeSciences, Rochester, MN), 2mM Glutamax (Life Technologies, Grand Island, NY), 2 U/mL heparin, and 1% Penn-Strep (100 U/mL penicillin, 100 μg/mL streptomycin; Cellgro, Corning, NY). All AMSCs used in experimentation were of passage 7.

Onishi K., Jones D.L., Riester S.M., Lewallen E.A., Lewallen D.G., Sellon J.L., Dietz A.B., Qu W., van Wijnen A.J, & Smith J. (2016). HUMAN ADIPOSE-DERIVED MESENCHYMAL STROMAL/STEM CELLS REMAIN VIABLE AND METABOLICALLY ACTIVE FOLLOWING NEEDLE PASSAGE. PM & R : the journal of injury, function, and rehabilitation, 8(9), 844-854.

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Isolation and Culture of MSCs from Liver and Adipose Tissue

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The study was approved by the Mayo Clinic Institutional Review Board. MSCs were isolated from liver tissue of deceased organ donors (L-MSC) and subcutaneous abdominal adipose tissue of living kidney donors and weight reduction surgery (A-MSC). These donors underwent screening and met the criteria for organ donation. All samples were processed within 8 h of procurement. Liver and adipose tissues were minced in a Petri dish and mixed with 0.075% Collagenase IV (STEMCELL Technologies, Cambridge, MA, United States) in Dulbecco’s phosphate-buffered saline. After 45 min, enzyme action was stopped by adding platelet lysate (PL5%) MSC media; Advanced Minimum Essential Medium (Thermo Fisher Scientific, Waltham, MA, United States), PLTGold Human Platelet Lysate (EMD Millipore, Burlington, MA, United States), and GlutaMAX (Thermo Fisher Scientific, Waltham, MA, United States). Digested tissue was centrifuged, resuspended in fresh media, and filtered twice before plating in PL5% MSC media. Non-adherent cells were removed every 3 days thereafter. Cultures were maintained at 37°C, 5% CO₂ in a humidified incubator.

Yigitbilek F., Conley S.M., Tang H., Saadiq I.M., Jordan K.L., Lerman L.O, & Taner T. (2021). Comparable in vitro Function of Human Liver-Derived and Adipose Tissue-Derived Mesenchymal Stromal Cells: Implications for Cell-Based Therapy. Frontiers in Cell and Developmental Biology, 9, 641792.

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Isolation of Knee Fibroblasts from Posterior Capsule

Cited 1 time

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Detailed descriptions of specimen collection, processing, and acquisition of primary knee outgrowth fibroblasts from the posterior capsule (PC) of three patients (i.e., 030, 256, and 565) undergoing primary TKA has been previously provided [Bayram et al., 2022 (link)]. Cells were grown in advanced minimum essential medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS) (R&D Systems, Minneapolis, MN), 1% Penicillin/Streptomycin (Thermo Fisher Scientific) and 1% Glutamax (Thermo Fisher Scientific) in cell culture incubators at 37°C, 95% humidity, 21% O2, and 5% CO2 conditions to establish outgrowth cell cultures. All experiments were performed using plastic tissue culture flasks and plates. Passage four and five knee outgrowth fibroblasts were used for all experiments, as was done in our recent study that defines the transcriptional profile and myofibroblast differentiation potential of these cells [Bayram et al., 2022 (link)].

Dudakovic A., Bayram B., Bettencourt J.W., Limberg A.K., Galvan M.L., Carrasco M.E., Stans B., Thaler R., Morrey M.E., Sanchez-Sotelo J., Berry D.J., van Wijnen A.J, & Abdel M.P. (2023). The Epigenetic Regulator BRD4 is Required for Myofibroblast Differentiation of Knee Fibroblasts. Journal of cellular biochemistry, 124(2), 320-334.

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Isolation and Characterization of Adipocyte Progenitors

Cited 1 time

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Adipocyte progenitors, also termed adipose tissue-derived mesenchymal stem cells or preadipocytes, were harvested from abdominal subcutaneous fat samples as previously described [48 (link),49 (link)]. Adipose biopsies were minced and digested in 2% collagenase type I at 37 °C for 45 min (Gibco, Waltham, MA), filtered through a 100 μm cell strainer (BD Biosciences, San José, CA) to remove remaining tissue pieces, and centrifuged to pellet cells. Cells were cultured in advanced minimum-essential-medium (Thermo Fisher Scientific, Waltham, MA) supplemented with 5% platelet lysate (PLTMax, Mill Creek Life Sciences, Rochester, MN) and 2 mM l-glutamine (Invitrogen, Carlsbad, CA) in a 37 °C/5% CO₂ incubator for 3–4 days. When 60–80% confluent, cells were passaged using TrypLE (Trypsin-like Enzyme, Invitrogen, Invitrogen, Waltham, MA). The third passage cells were collected and kept in Gibco Cell Culture Freezing Medium (Life Technologies, Carlsbad, CA) at −80 °C. Change in adipocyte progenitor numbers over time was measured in passage 3–4 cultures using a tetrazolium compound (3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium, inner salt; MTS) assay (CellTiter 96 Non-Radioactive Cell Proliferation Assay; Promega, Madison, WI) according to the manufacturer's instructions.

Hickson L.J., Langhi Prata L.G., Bobart S.A., Evans T.K., Giorgadze N., Hashmi S.K., Herrmann S.M., Jensen M.D., Jia Q., Jordan K.L., Kellogg T.A., Khosla S., Koerber D.M., Lagnado A.B., Lawson D.K., LeBrasseur N.K., Lerman L.O., McDonald K.M., McKenzie T.J., Passos J.F., Pignolo R.J., Pirtskhalava T., Saadiq I.M., Schaefer K.K., Textor S.C., Victorelli S.G., Volkman T.L., Xue A., Wentworth M.A., Wissler Gerdes E.O., Zhu Y., Tchkonia T, & Kirkland J.L. (2019). Senolytics decrease senescent cells in humans: Preliminary report from a clinical trial of Dasatinib plus Quercetin in individuals with diabetic kidney disease. EBioMedicine, 47, 446-456.

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