Matrigel is a commercially available substrate that is derived from the extracellular matrix. Matrigel is widely used in cell culture experiments such as the transdifferentiation of primary pancreatic acini to ductal epithelial-like cells. Difficulty arises during gene expression analysis for cells cultured on Matrigel because residual RNA in the Matrigel will not only contribute to the poor integrity of RNA isolated from Matrigel cultures, but also will impact the gene expression data. We report here a simple method of removing Matrigel from primary cultures of human or mouse pancreatic acini. Following the experiment, the cultures are placed on wet ice to liquefy the Matrigel. The cell and Matrigel mixture is then centrifuged at low speed to separate the pancreatic cells from the Matrigel solution that resides in the supernatant. RNA isolated from the pelleted cells has high integrity and may be readily used for gene expression analysis such as quantitative reverse transcription PCR. (C) 2020 The Authors. Published by Elsevier B.V.
机构:
Univ Michigan, Program Cellular & Mol Biol, Ann Arbor, MI 48109 USAUniv Michigan, Program Cellular & Mol Biol, Ann Arbor, MI 48109 USA
Collins, Meredith A.
;
Yan, Wei
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Univ Michigan, Dept Pathol, Ann Arbor, MI 48109 USA
Univ Michigan, Michigan Ctr Translat Pathol, Ann Arbor, MI 48109 USAUniv Michigan, Program Cellular & Mol Biol, Ann Arbor, MI 48109 USA
Yan, Wei
;
Sebolt-Leopold, Judith S.
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Univ Michigan, Dept Radiol, Ann Arbor, MI 48109 USAUniv Michigan, Program Cellular & Mol Biol, Ann Arbor, MI 48109 USA
Sebolt-Leopold, Judith S.
;
di Magliano, Marina Pasca
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Univ Michigan, Program Cellular & Mol Biol, Ann Arbor, MI 48109 USA
Univ Michigan, Dept Surg, Ann Arbor, MI 48109 USA
Univ Michigan, Dept Cell & Dev Biol, Ann Arbor, MI 48109 USAUniv Michigan, Program Cellular & Mol Biol, Ann Arbor, MI 48109 USA
机构:
New Mexico State Univ, Mol Biol Program, Las Cruces, NM 88003 USA
Suny Downstate Med Ctr, Dept Pediat, Brooklyn, NY 11203 USA
Suny Downstate Med Ctr, Dept Cell Biol, Brooklyn, NY 11203 USANew Mexico State Univ, Mol Biol Program, Las Cruces, NM 88003 USA
Jogalekar, Manasi P.
;
Serrano, Elba E.
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New Mexico State Univ, Dept Biol, Las Cruces, NM 88003 USANew Mexico State Univ, Mol Biol Program, Las Cruces, NM 88003 USA
机构:
Univ Michigan, Program Cellular & Mol Biol, Ann Arbor, MI 48109 USAUniv Michigan, Program Cellular & Mol Biol, Ann Arbor, MI 48109 USA
Collins, Meredith A.
;
Yan, Wei
论文数: 0引用数: 0
h-index: 0
机构:
Univ Michigan, Dept Pathol, Ann Arbor, MI 48109 USA
Univ Michigan, Michigan Ctr Translat Pathol, Ann Arbor, MI 48109 USAUniv Michigan, Program Cellular & Mol Biol, Ann Arbor, MI 48109 USA
Yan, Wei
;
Sebolt-Leopold, Judith S.
论文数: 0引用数: 0
h-index: 0
机构:
Univ Michigan, Dept Radiol, Ann Arbor, MI 48109 USAUniv Michigan, Program Cellular & Mol Biol, Ann Arbor, MI 48109 USA
Sebolt-Leopold, Judith S.
;
di Magliano, Marina Pasca
论文数: 0引用数: 0
h-index: 0
机构:
Univ Michigan, Program Cellular & Mol Biol, Ann Arbor, MI 48109 USA
Univ Michigan, Dept Surg, Ann Arbor, MI 48109 USA
Univ Michigan, Dept Cell & Dev Biol, Ann Arbor, MI 48109 USAUniv Michigan, Program Cellular & Mol Biol, Ann Arbor, MI 48109 USA
机构:
New Mexico State Univ, Mol Biol Program, Las Cruces, NM 88003 USA
Suny Downstate Med Ctr, Dept Pediat, Brooklyn, NY 11203 USA
Suny Downstate Med Ctr, Dept Cell Biol, Brooklyn, NY 11203 USANew Mexico State Univ, Mol Biol Program, Las Cruces, NM 88003 USA
Jogalekar, Manasi P.
;
Serrano, Elba E.
论文数: 0引用数: 0
h-index: 0
机构:
New Mexico State Univ, Dept Biol, Las Cruces, NM 88003 USANew Mexico State Univ, Mol Biol Program, Las Cruces, NM 88003 USA