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Amniomax c 100 complete medium

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The AmnioMAX C-100 complete medium is a cell culture medium designed for the growth and maintenance of human amniotic fluid cells. It provides the necessary nutrients and growth factors to support the optimal proliferation and viability of these cell types in vitro.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions) Gentamycine, by Thermo Fisher Scientific (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) Staurosporine, by Alexis Biochemicals (2 mentions) Dharmafect 1, by Thermo Fisher Scientific (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Mg132, by Cayman Chemical (2 mentions) Amv reverse transcriptase, by Roche (2 mentions) Rneasy kit, by Qiagen (2 mentions) Rnaimax, by Thermo Fisher Scientific (2 mentions) Dmem f12, by Thermo Fisher Scientific (2 mentions) Aphidicolin, by Merck Group (1 mentions) Icrf 159, by Merck Group (1 mentions) Neon transfection system, by Thermo Fisher Scientific (1 mentions) E gel electrophoresis system, by Thermo Fisher Scientific (1 mentions) Dmem 10 fbs, by Merck Group (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Amniomax C-100 medium Amniomax C-100 AmnioMax medium Complete medium AmnioMAX

12 protocols using amniomax c 100 complete medium

1

Tumor Tissue Dissociation and Cell Culture

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Tumor pieces were collected into DMEM (Dulbecco’s Modified Eagles Medium, PAA Laboratories GmbH, Pasching, Austria). Biopsies were cut into 1 mm3 pieces and placed into either DMEM for immediate processing or into freezing medium (90% serum, 10% DMSO) prior to being progressively cooled to −80°C. Cell dissociation was performed mechanically by passage through increasingly finer needles (19G to 26G). Single cells were seeded in AmnioMAX™ C-100 complete medium containing gentamycine, L-glutamine and FBS (Gibco, Invitrogen, Paisley, UK) and maintained at 37°C in a 5% CO2 humidified atmosphere. The cells were further cultured until appearance of adherent cells and colony formation and then weekly passaged.
Grasso C.S., Tang Y., Truffaux N., Berlow N.E., Liu L., Debily M.A., Quist M.J., Davis L.E., Huang E.C., Woo P.J., Ponnuswami A., Chen S., Johung T.B., Sun W., Kogiso M., Du Y., Lin Q., Huang Y., Hütt-Cabezas M., Warren K.E., Dret L.L., Meltzer P.S., Mao H., Quezado M., van Vuurden D.G., Abraham J., Fouladi M., Svalina M.N., Wang N., Hawkins C., Nazarian J., Alonso M.M., Raabe E., Hulleman E., Spellman P.T., Li X.N., Keller C., Pal R., Grill J, & Monje M. (2015). Functionally-defined Therapeutic Targets in Diffuse Intrinsic Pontine Glioma: A Report of the Children’s Oncology Group DIPG Preclinical Consortium. Nature medicine, 21(6), 555-559.
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2

Tumor Tissue Dissociation and Cell Culture

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Tumor pieces were collected into DMEM (Dulbecco’s Modified Eagles Medium, PAA Laboratories GmbH, Pasching, Austria). Biopsies were cut into 1 mm3 pieces and placed into either DMEM for immediate processing or into freezing medium (90% serum, 10% DMSO) prior to being progressively cooled to −80°C. Cell dissociation was performed mechanically by passage through increasingly finer needles (19G to 26G). Single cells were seeded in AmnioMAX™ C-100 complete medium containing gentamycine, L-glutamine and FBS (Gibco, Invitrogen, Paisley, UK) and maintained at 37°C in a 5% CO2 humidified atmosphere. The cells were further cultured until appearance of adherent cells and colony formation and then weekly passaged.
Grasso C.S., Tang Y., Truffaux N., Berlow N.E., Liu L., Debily M.A., Quist M.J., Davis L.E., Huang E.C., Woo P.J., Ponnuswami A., Chen S., Johung T.B., Sun W., Kogiso M., Du Y., Lin Q., Huang Y., Hütt-Cabezas M., Warren K.E., Dret L.L., Meltzer P.S., Mao H., Quezado M., van Vuurden D.G., Abraham J., Fouladi M., Svalina M.N., Wang N., Hawkins C., Nazarian J., Alonso M.M., Raabe E., Hulleman E., Spellman P.T., Li X.N., Keller C., Pal R., Grill J, & Monje M. (2015). Functionally-defined Therapeutic Targets in Diffuse Intrinsic Pontine Glioma: A Report of the Children’s Oncology Group DIPG Preclinical Consortium. Nature medicine, 21(6), 555-559.
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3

Ciliogenesis Induction in Dermal Fibroblasts

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Dermal fibroblasts were obtained by skin punch biopsy and were cultured in amnioMAX C-100 complete medium (Life Technologies) and maintained in a 37°C incubator with 5% CO2 and 3% O2. HeLa were obtained from European collection of cell cultures (#93021013) cultured in DMEM (Life technologies), 10% Fetal Calf Serum (FCS), 100 U/ml penicillin and 100 μg/ml streptomycin. All cell lines were routinely tested for mycoplasma. To induce ciliogenesis, cells were incubated in low serum (0.5% FCS) DMEM medium for 48 hrs.
Short interfering RNA (siRNA) oligonucleotides were transfected into primary fibroblasts using Dharmafect 1 (Thermo Fischer) according to manufacturer’s instructions. Oligonucleotide sequences used are in Table S3e. PLK4 expression vectors were transfected into HeLa cells using lipofectamine 2000 (Life technologies) according to manufacturer’s instructions. Where indicated, cells were treated with 10 μM MG132 (Cayman Chemicals) or 1.5 μM staurosporine (Alexis Biochemicals) for 5 hrs.
Martin C.A., Ahmad I., Klingseisen A., Hussain M.S., Bicknell L.S., Leitch A., Nürnberg G., Toliat M.R., Murray J.E., Hunt D., Khan F., Ali Z., Tinschert S., Ding J., Keith C., Harley M.E., Heyn P., Müller R., Hoffmann I., Cormier-Daire V., Dollfus H., Dupuis L., Bashamboo A., McElreavey K., Kariminejad A., Mendoza-Londono R., Moore A.T., Saggar A., Schlechter C., Weleber R., Thiele H., Altmüller J., Höhne W., Hurles M.E., Noegel A.A., Baig S.M., Nürnberg P, & Jackson A.P. (2014). Mutations in PLK4, encoding a master regulator of centriole biogenesis, cause microcephaly, growth failure and retinopathy. Nature genetics, 46(12), 1283-1292.
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4

Fibroblast-based PUF60 Allele Sequencing

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Dermal fibroblasts were obtained from patient 8 (3781-3781) by skin punch biopsy and cultured in amnioMAX C-100 complete medium (Life Technologies) as described previously12 (link). RNA was extracted from primary skin-derived fibroblast cell lines from patient 8 (3781-3781) and two sex-matched controls using RNeasy kit (Qiagen) and treated with DNAseI to eliminate genomic DNA, according to manufacturer’s instructions. Complementary DNA (cDNA) synthesis was carried out using random oligomer primers and AMV Reverse Transcriptase (Roche). The cDNA samples were resolved on an E-Gel® electrophoresis system and extracted according to manufacturer’s instructions (ThermoFisher Scientific) in order to sequence the amplicons corresponding to the normal and mutant alleles. Sequences of primers used for cDNA amplification and Sanger sequencing of PUF60 are available upon request.
Low K.J., Ansari M., Abou Jamra R., Clarke A., El Chehadeh S., FitzPatrick D.R., Greenslade M., Henderson A., Hurst J., Keller K., Kuentz P., Prescott T., Roessler F., Selmer K.K., Schneider M.C., Stewart F., Tatton-Brown K., Thevenon J., Vigeland M.D., Vogt J., Willems M., Zonana J., Study D.D, & Smithson S.F. (2017). PUF60 variants cause a syndrome of ID, short stature, microcephaly, coloboma, craniofacial, cardiac, renal and spinal features. European Journal of Human Genetics, 25(5), 552-559.
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5

Isolation of Chorionic Mesenchymal Stem Cells

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CMSCs were isolated using the explant method as described previously [7 (link)] with the following modifications. Briefly, an incision was made through the fetal membranes near the umbilical cord insertion site and 1 g of chorionic villous tissue was obtained from approximately 1–2 cm below the chorionic plate. Pieces of chorionic tissue with typical villous morphology were cleaned with a 21 gauge needle under a dissecting microscope to remove non-villous tissue. Cleaned villi were finely diced and digested in 0.25% trypsin for 40 min at 37°C. The trypsin was inactivated with FBS and tissues were washed in PBS. The digested villi were cultured in Amniomax C100 complete medium (Life Technologies) in 25 cm2 tissue culture flasks maintained at 37°C in a humidified 5% CO2 incubator. After 7 days, villous tissues were removed from the flask and the adherent cells arising from the explants (P0 cells) were grown until at least 80% confluent before expanding to reach P5.
Kusuma G.D., Menicanin D., Gronthos S., Manuelpillai U., Abumaree M.H., Pertile M.D., Brennecke S.P, & Kalionis B. (2015). Ectopic Bone Formation by Mesenchymal Stem Cells Derived from Human Term Placenta and the Decidua. PLoS ONE, 10(10), e0141246.
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6

Dermal Fibroblast Culture and siRNA Transfection

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Dermal fibroblasts were obtained by skin-punch biopsy and were maintained in amnioMAX C-100 complete medium (Life Technologies) in a 37°C incubator with 5% CO2 and 3% O2. siRNA oligonucleotides (siLUC: 5′-CUUACGCUGAGUACUUCGA-3′ [siTopIIIα SMARTpool M-005279-01-0005, Dharmacon]) were transfected into dermal fibroblasts with RNAiMAX (Life Technologies) according to the manufacturer’s instructions.
Martin C.A., Sarlós K., Logan C.V., Thakur R.S., Parry D.A., Bizard A.H., Leitch A., Cleal L., Ali N.S., Al-Owain M.A., Allen W., Altmüller J., Aza-Carmona M., Barakat B.A., Barraza-García J., Begtrup A., Bogliolo M., Cho M.T., Cruz-Rojo J., Dhahrabi H.A., Elcioglu N.H., Gorman G.S., Jobling R., Kesterton I., Kishita Y., Kohda M., Le Quesne Stabej P., Malallah A.J., Nürnberg P., Ohtake A., Okazaki Y., Pujol R., Ramirez M.J., Revah-Politi A., Shimura M., Stevens P., Taylor R.W., Turner L., Williams H., Wilson C., Yigit G., Zahavich L., Alkuraya F.S., Surralles J., Iglesais A., Murayama K., Wollnik B., Dattani M., Heath K.E., Hickson I.D, & Jackson A.P. (2018). Mutations in TOP3A Cause a Bloom Syndrome-like Disorder. American Journal of Human Genetics, 103(2), 221-231.
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7

Culturing and Transfecting Fibroblast Cell Lines

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Dermal fibroblasts were obtained by skin punch biopsy and cultured in amnioMAX C-100 complete medium (Life Technologies). MEFs were prepared from individual E13.5 embryos after removing the head and abdominal cavities. Each embryo was minced and maintained in DMEM, 10% FCS, 0.1 mM β-mercaptoethanol, 100 U/mL penicillin, and 100 µg/mL streptomycin. All fibroblast cultures were maintained in a 37°C incubator with 5% CO2 and 3% O2. hTERT RPE-1 cells were obtained from the American Type Culture Collection (CRL-4000) and cultured in DMEM:F12, 10% FCS, 0.26% sodium bicarbonate, 100 U/mL penicillin, and 100 µg/mL streptomycin. All cell lines were routinely tested for mycoplasma. siRNA oligonucleotides were transfected into hTERT RPE1 cells using RNAiMAX (Life Technologies) according to the manufacturer's instructions. For siRNA oligonucleotide sequences, see Supplemental Table S2C. pRFP-H2B was transfected into primary fibroblast cells by electroporation with the Neon transfection system (Thermo Fischer Scientific) according to the manufacturer's instructions. Where indicated, cells were treated with 0.3 µM aphidicolin (Sigma-Aldrich) or 5 µM ICRF-159 (Sigma-Aldrich) for 24 h.
Martin C.A., Murray J.E., Carroll P., Leitch A., Mackenzie K.J., Halachev M., Fetit A.E., Keith C., Bicknell L.S., Fluteau A., Gautier P., Hall E.A., Joss S., Soares G., Silva J., Bober M.B., Duker A., Wise C.A., Quigley A.J., Phadke S.R., Wood A.J., Vagnarelli P, & Jackson A.P. (2016). Mutations in genes encoding condensin complex proteins cause microcephaly through decatenation failure at mitosis. Genes & Development, 30(19), 2158-2172.
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8

Fibroblast Transcriptome Analysis of Genetic Disorder

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Dermal fibroblasts were obtained from patient 8 (3781–3781) by skin punch biopsy and cultured in amnioMAX C-100 complete medium (Life Technologies, Carlsbad, CA, USA) as described previously.12 (link) RNA was extracted from primary skin-derived fibroblast cell lines from patient 8 (3781–3781) and two sex-matched controls using RNeasy Kit (Qiagen, Hilden, Germany) and treated with DNAseI to eliminate genomic DNA, according to the manufacturer's instructions. Complementary DNA (cDNA) synthesis was carried out using random oligomer primers and AMV Reverse Transcriptase (Roche, Penzburg, Germany). The cDNA samples were resolved on an E-Gel electrophoresis system and extracted according to the manufacturer's instructions (Thermo Fisher Scientific, Waltham, MA, USA) to sequence the amplicons corresponding to the normal and mutant alleles. Sequences of primers used for cDNA amplification and Sanger sequencing of PUF60 are available upon request.
Low K.J., Ansari M., Abou Jamra R., Clarke A., El Chehadeh S., FitzPatrick D.R., Greenslade M., Henderson A., Hurst J., Keller K., Kuentz P., Prescott T., Roessler F., Selmer K.K., Schneider M.C., Stewart F., Tatton-Brown K., Thevenon J., Vigeland M.D., Vogt J., Willems M., Zonana J., Study D.D, & Smithson S.F. (2017). PUF60 variants cause a syndrome of ID, short stature, microcephaly, coloboma, craniofacial, cardiac, renal and spinal features. European Journal of Human Genetics, 25(5), 552-559.
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9

Ciliogenesis Induction in Dermal Fibroblasts

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Dermal fibroblasts were obtained by skin punch biopsy and were cultured in amnioMAX C-100 complete medium (Life Technologies) and maintained in a 37°C incubator with 5% CO2 and 3% O2. HeLa were obtained from European collection of cell cultures (#93021013) cultured in DMEM (Life technologies), 10% Fetal Calf Serum (FCS), 100 U/ml penicillin and 100 μg/ml streptomycin. All cell lines were routinely tested for mycoplasma. To induce ciliogenesis, cells were incubated in low serum (0.5% FCS) DMEM medium for 48 hrs.
Short interfering RNA (siRNA) oligonucleotides were transfected into primary fibroblasts using Dharmafect 1 (Thermo Fischer) according to manufacturer’s instructions. Oligonucleotide sequences used are in Table S3e. PLK4 expression vectors were transfected into HeLa cells using lipofectamine 2000 (Life technologies) according to manufacturer’s instructions. Where indicated, cells were treated with 10 μM MG132 (Cayman Chemicals) or 1.5 μM staurosporine (Alexis Biochemicals) for 5 hrs.
Martin C.A., Ahmad I., Klingseisen A., Hussain M.S., Bicknell L.S., Leitch A., Nürnberg G., Toliat M.R., Murray J.E., Hunt D., Khan F., Ali Z., Tinschert S., Ding J., Keith C., Harley M.E., Heyn P., Müller R., Hoffmann I., Cormier-Daire V., Dollfus H., Dupuis L., Bashamboo A., McElreavey K., Kariminejad A., Mendoza-Londono R., Moore A.T., Saggar A., Schlechter C., Weleber R., Thiele H., Altmüller J., Höhne W., Hurles M.E., Noegel A.A., Baig S.M., Nürnberg P, & Jackson A.P. (2014). Mutations in PLK4, encoding a master regulator of centriole biogenesis, cause microcephaly, growth failure and retinopathy. Nature genetics, 46(12), 1283-1292.
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10

Isolation and Culture of Uterine Leiomyoma Cells

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Human pancreatic (PANC-1) and human kidney (HEK293T) cells were purchased from the Cell Collection of Institute of Cytology RAS (Saint-Petersburg, Russia). The cells were cultured according to the standard method “Fundamental Techniques in Cell Culture” SIGMA-ALDRICH (Sailsbury Wiltshire, SP4, 0JG, UK). Primary UL cells were isolated from myomatous nodes after hysterectomy was carried out in the D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology (Saint-Petersburg, Russia), as described previously [52 (link)]. Briefly, the UL cells were propagated in AmnioMAX C-100 Complete Medium (Thermo Fisher Scientific, Carlsbad, CA, USA) supplemented with 10% FBS (Thermo Fisher Scientific) and 0.3% gentamicin. Only p1 cell culture was used for the transfection experiments.
Egorova A., Selutin A., Maretina M., Selkov S, & Kiselev A. (2022). Peptide-Based Nanoparticles for αvβ3 Integrin-Targeted DNA Delivery to Cancer and Uterine Leiomyoma Cells. Molecules, 27(23), 8363.
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