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Miami media 1a

Manufactured by Corning
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Sourced in United States

The Miami Media #1A is a compact and versatile laboratory equipment designed for media preparation. It features a digital control panel, temperature regulation, and a secure lid to maintain sterile conditions during operation. The core function of this product is to facilitate the efficient preparation of culture media for various scientific applications.

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Lab products found in correlation

Final wash culture medium, by Corning (2 mentions) Tazemetostat, by Selleck Chemicals (1 mentions) Gsk126, by Selleck Chemicals (1 mentions)

14 protocols using miami media 1a

1

Islet Stimulation and Immunostaining

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Human islets were obtained from the Integrated Islet Distribution Program. All studies and protocols used were approved by the Joslin Diabetes Center’s Committee on Human Studies (CHS#5-05). Upon receipt, islets were cultured overnight in Miami Media #1A (Cellgro). The islets were then starved in Final Wash/Culture Media (Cellgro) for 3-hours before being stimulated with Miami Media #1A supplemented with sivelestat or GW311616A. Twenty four hrs later, islets were embedded in agarose and used for immunostaining studies (described below).
El Ouaamari A., Dirice E., Gedeon N., Hu J., Zhou J.Y., Shirakawa J., Hou L., Goodman J., Karampelias C., Qiang G., Boucher J., Martinez R., Gritsenko M.A., De Jesus D.F., Kahraman S., Bhatt S., Smith R.D., Beer H.D., Jungtrakoon P., Gong Y., Goldfine A.B., Liew C.W., Doria A., Andersson O., Qian W.J., Remold-O'Donnell E, & Kulkarni R.N. (2015). SerpinB1 Promotes Pancreatic β-Cell Proliferation. Cell metabolism, 23(1), 194-205.
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2

Isolation and Purification of Human Islets

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Human islets were obtained from the Integrated Islet Distribution Program (IIDP) and Prodo Laboratories. Upon receipt, islets were cultured overnight (16h) in Miami Media #1A (Cellgro, USA). Islets were then handpicked, washed 2 times by self-sedimentation with ice-cold Dulbecco’s phosphate buffered saline (DPBS) and 20,000 islets equivalents (IE) were pelleted for RNA isolation.
De Jesus D.F., Zhang Z., Kahraman S., Brown N.K., Chen M., Hu J., Gupta M.K., He C, & Kulkarni R.N. (2019). m6A mRNA Methylation Regulates Human β-Cell Biology in Physiological States and in Type 2 Diabetes. Nature metabolism, 1(8), 765-774.
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3

Serum-Stimulated Human Islet Analysis

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Human islets were obtained from Clinical Islet Laboratory and Clinical Islet Transplant Program of University of Alberta or the Alberta Diabetes Institute Islet Core of the University of Alberta. All studies and protocols used were approved by Yokohama City University Ethics Board (approval B171100025) and the Joslin Diabetes Center’s Committee on Human Studies (approval CHS#5–05). Details of human islets are described in the ESM Human Islet Checklist. Human islets were handpicked and cultured in Miami Media #1A (Cellgro, Mediatech, Manassas, VA, USA). For the stimulation with serum, the islets were cultured in Final Wash/Culture Medium (Cellgro) containing 20% serum (vol./vol.) from control, Luse-, OSI-906- or OSI-906+Luse-treated mice treated for 48 h in the presence of 5-ethynyl-2-deoxyuridine (EdU). The sera were collected from mice at day 7 of treatment. The islets were embedded in agarose and used for immunostaining studies.
Shirakawa J., Tajima K., Okuyama T., Kyohara M., Togashi Y., De Jesus D.F., Basile G., Kin T., Shapiro A.M., Kulkarni R.N, & Terauchi Y. (2020). Luseogliflozin increases beta cell proliferation through humoral factors that activate an insulin receptor- and IGF-1 receptor-independent pathway. Diabetologia, 63(3), 577-587.
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4

Isolation and Purification of Human Islets

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Human islets were obtained from the Integrated Islet Distribution Program (IIDP) and Prodo Laboratories. Upon receipt, islets were cultured overnight (16h) in Miami Media #1A (Cellgro, USA). Islets were then handpicked, washed 2 times by self-sedimentation with ice-cold Dulbecco’s phosphate buffered saline (DPBS) and 20,000 islets equivalents (IE) were pelleted for RNA isolation.
De Jesus D.F., Zhang Z., Kahraman S., Brown N.K., Chen M., Hu J., Gupta M.K., He C, & Kulkarni R.N. (2019). m6A mRNA Methylation Regulates Human β-Cell Biology in Physiological States and in Type 2 Diabetes. Nature metabolism, 1(8), 765-774.
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5

Methodology for Insulin Stimulation of Human Islets

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Human islets were obtained from the Integrated Islet Distribution Program. Details of human islets are described in Table S4. All studies and protocols used were approved by the Joslin Diabetes Center’s Committee on Human Studies (CHS#5-05). Upon receipt, islets were cultured overnight in Miami Media #1A (Cellgro). The islets were embedded in agarose and used for immunostaining studies. For the stimulation with insulin, the islets were starved in Final Wash/Culture Medium (Cellgro) for 5-hours before being stimulated with 10 nM insulin.
Shirakawa J., Fernandez M., Takatani T., El Ouaamari A., Jungtrakoon P., Okawa E.R., Zhang W., Yi P., Doria A, & Kulkarni R.N. (2017). Insulin signaling regulates the FoxM1/PLK1/CENP-A pathway to promote adaptive pancreatic β-cell proliferation. Cell metabolism, 25(4), 868-882.e5.
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6

Islets Sourcing and Preparation

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A minimum of 20,000 human islets equivalents (IEQs)/patient were obtained from the Integrated Islet Distribution Program (IIIDP) and Prodo Laboratories. Upon receipt, islets were cultured overnight in Miami Media #1A (Cellgro, USA) and then handpicked, washed twice by self-sedimentation with ice-cold DPBS, and pelleted for RNA isolation. All studies and protocols used were approved by the Joslin Diabetes Center’s Committee on Human Studies (CHS#5-05). Samples from eight T2D patients and seven non-diabetic controls were collected for analyses in this study.
Zhang Z., Zhan Q., Eckert M., Zhu A., Chryplewicz A., De Jesus D.F., Ren D., Kulkarni R.N., Lengyel E., He C, & Chen M. (2019). RADAR: differential analysis of MeRIP-seq data with a random effect model. Genome Biology, 20, 294.
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7

Human Islet Isolation and Culture

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Human islets (3 males and 4 females, 22–52 years old) were obtained from the Clinical Islet Laboratory and Clinical Islet Transplant Program of University of Alberta or the Alberta Diabetes Institute IsletCore of the University of Alberta. Ethics approval and informed consent from donors or families were obtained in each institute. Details of human islets are described in Table S2. All studies and protocols used were approved by Yokohama City University Ethics Board (approval B171100025) and the Joslin Diabetes Center’s Committee on Human Studies (approval CHS#5–05). Upon receipt, islets were cultured overnight in Miami Media #1A (Cellgro). The islets were embedded in agarose and used for immunostaining studies.
Shirakawa J., Togashi Y., Basile G., Okuyama T., Inoue R., Fernandez M., Kyohara M., De Jesus D.F., Goto N., Zhang W., Tsuno T., Kin T., Pan H., Dreyfuss J.M., Shapiro A.M., Yi P., Terauchi Y, & Kulkarni R.N. (2022). E2F1 transcription factor mediates a link between fat and islets to promote β cell proliferation in response to acute insulin resistance. Cell reports, 41(1), 111436.
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8

Pregnancy Effects on Human Islet Grafts

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Upon receipt, human islets and human ductal aggregates isolated from nondiabetic donors (n = 4) were cultured overnight in Miami Media 1A (Cellgro). Hand-picked and size-matched islets (1000 IEQ) were transplanted together with 100 human ductal aggregates (cluster of ductal cells) under the kidney capsule of both NSG-Lox and NSG-LIRKO mice as described previously (56 (link), 57 (link)). After allowing 10 days after transplantation for islet engraftment, mice were either maintained in a nonpregnant state or allowed to breed to become pregnant. Human islet and duct grafts were removed on pregnancy day 15.5 and snap-frozen for further analysis.
Dirice E., Basile G., Kahraman S., Diegisser D., Hu J, & Kulkarni R.N. (2022). Single-nucleus RNA-Seq reveals singular gene signatures of human ductal cells during adaptation to insulin resistance. JCI Insight, 7(16), e153877.
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9

Isolation and Culture of Human Pancreatic Islets

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Human islets were obtained from 5 non-diabetic brain-dead donors. Islet preparations were generated by the Integrated Islet Distribution Program or Prodo laboratories according to the standard procedures [50 (link)] (Additional file 1: Table S1). All studies and protocols used were approved by the Joslin Diabetes Center’s Committee on Human Studies (CHS#5-05). Upon receipt, human islets were centrifuged, washed, transferred to Petri dishes (2000–3000 human islets equivalents [IEQs]/plate), and cultured with fresh Miami Media #1A (Cellgro) overnight at 37 °C and 5% CO2. Human islets from one donor (N = 1) were processed to isolate nuclei and obtain single-cell preparations as a common source for the scRNA-seq and snRNA-seq procedures (detailed below), while human islets from 4 donors (N = 4) were used in the kidney capsule transplantation experiments (detailed below).
Basile G., Kahraman S., Dirice E., Pan H., Dreyfuss J.M, & Kulkarni R.N. (2021). Using single-nucleus RNA-sequencing to interrogate transcriptomic profiles of archived human pancreatic islets. Genome Medicine, 13, 128.
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10

Human Islet Isolation and Culture

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Human islets (3 males and 4 females, 22–52 years old) were obtained from the Clinical Islet Laboratory and Clinical Islet Transplant Program of University of Alberta or the Alberta Diabetes Institute IsletCore of the University of Alberta. Ethics approval and informed consent from donors or families were obtained in each institute. Details of human islets are described in Table S2. All studies and protocols used were approved by Yokohama City University Ethics Board (approval B171100025) and the Joslin Diabetes Center’s Committee on Human Studies (approval CHS#5–05). Upon receipt, islets were cultured overnight in Miami Media #1A (Cellgro). The islets were embedded in agarose and used for immunostaining studies.
Shirakawa J., Togashi Y., Basile G., Okuyama T., Inoue R., Fernandez M., Kyohara M., De Jesus D.F., Goto N., Zhang W., Tsuno T., Kin T., Pan H., Dreyfuss J.M., Shapiro A.M., Yi P., Terauchi Y, & Kulkarni R.N. (2022). E2F1 transcription factor mediates a link between fat and islets to promote β cell proliferation in response to acute insulin resistance. Cell reports, 41(1), 111436.
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