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Matrigel solubilized basement membrane

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Matrigel solubilized basement membrane is a gelatinous protein mixture extracted from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells. It is composed of laminin, collagen IV, heparan sulfate proteoglycans, and various growth factors. The core function of Matrigel is to provide a complex and biologically active extracellular matrix (ECM) that supports the growth and differentiation of cells in in vitro cell culture systems.

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Lab products found in correlation

Sustained release testosterone pellets, by Innovative Research (3 mentions) Cs12708, by Cell Signaling Technology (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Matrigel basement membrane Basement Matrigel Matrigel membrane Matrigel

5 protocols using matrigel solubilized basement membrane

1

Xenograft Tumor Model for Prostate Cancer

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Mice (4–5-week-old, nu/nu athymic male) were obtained from Harlan Sprague-Dawley, Inc. and implanted subcutaneously (s.c.) with sustained release testosterone pellets (12.5mg, 90-day release; Innovative Research of America). All animal experiments were carried out in accordance with a protocol approved by the UC Davis Institutional Animal Care and Use Committee (IACUC). Suspensions of CWR22 (kindly provided by Dr. Clifford Tepper, Department of Biochemistry, UC Davis) or 22Rv1 cells in 50% Matrigel solubilized basement membrane (BD Biosciences), and tumors were established by s.c. injections of 2.5×106 cells/site into both flanks. Tumor-bearing mice were left intact or castrated by standard procedures (45). When palpable tumors were observed, animals were randomly assigned to one of the following groups (n=6 per group) and were treated with (a) vehicle (peanut oil only), (b) bicalutamide, delivered by oral gavage at a dose of 50mg/Kg, 100μL per dose, five-times per week, dissolved in ethanol, and delivered as a suspension in peanut oil, (c) rapamycin at a dose of 4mg/Kg 5-times a week by oral gavage, delivered as a suspension in peanut oil and (d) bicalutamide+rapamycin. At the end, mice were euthanized; and tumors collected for paraffin embedding. Blood from the euthanized animals was fractionated and serum collected for analysis.
D’Abronzo L.S., Bose S., Crapuchettes M.E., Beggs R.E., Vinall R.L., Tepper C.G., Siddiqui S., Mudryj M., Melgoza F.U., Durbin-Johnson B.P., deVere White R.W, & Ghosh. P.M. (2017). The Androgen Receptor is a negative regulator of eIF4E Phosphorylation at S209: Implications for the use of mTOR inhibitors in advanced prostate cancer. Oncogene, 36(46), 6359-6373.
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2

Xenograft Tumor Establishment and Immunohistochemistry

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All animal studies were conducted, and data collected with approval from the University of California Davis institutional animal care and use committee. Animal studies were performed as described previously (29 (link)). In summary, 4- to 5-week-old male Balb/c athymic Foxn1nu (nu/nu) mice (Harlan Sprague Dawley, Inc) were subcutaneously implanted in the flank with suspensions of CWR22 or 22Rv1 (2.5 million cells/site) in 50% Matrigel-solubilized basement membrane (BD Biosciences) and xenografts established. When palpable tumors were observed, the animals were castrated by bilateral scrotal excision, following isoflurane anesthetization or sham-operated by opening the animals surgically, but no tissues were removed. Mice were sacrificed when tumor size exceeded 150 mm in any one dimension or at the end of the study period. Mouse tumors were fixed in 10% buffered formalin (Medical Industries) for 30 min at RT, after which the pellet was immersed in 600-μl liquefied agar at 50 to 60 °C. The agar containing the tumor was paraffin-embedded and processed based on established protocols. Tumors were then stained with rabbit polyclonal anti-ErbB3 (CS-12708; 1:100 dilution, Cell Signaling Technologies).
Jathal M.K., Siddiqui S., Vasilatis D.M., Durbin Johnson B.P., Drake C., Mooso B.A., D’Abronzo L.S., Batra N., Mudryj M, & Ghosh P.M. (2023). Androgen receptor transcriptional activity is required for heregulin-1β–mediated nuclear localization of the HER3/ErbB3 receptor tyrosine kinase. The Journal of Biological Chemistry, 299(8), 104973.
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3

Xenograft Tumor Model for Prostate Cancer

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Mice (4–5-week-old, nu/nu athymic male) were obtained from Harlan Sprague-Dawley, Inc. and implanted subcutaneously (s.c.) with sustained release testosterone pellets (12.5mg, 90-day release; Innovative Research of America). All animal experiments were carried out in accordance with a protocol approved by the UC Davis Institutional Animal Care and Use Committee (IACUC). Suspensions of CWR22 (kindly provided by Dr. Clifford Tepper, Department of Biochemistry, UC Davis) or 22Rv1 cells in 50% Matrigel solubilized basement membrane (BD Biosciences), and tumors were established by s.c. injections of 2.5×106 cells/site into both flanks. Tumor-bearing mice were left intact or castrated by standard procedures (45). When palpable tumors were observed, animals were randomly assigned to one of the following groups (n=6 per group) and were treated with (a) vehicle (peanut oil only), (b) bicalutamide, delivered by oral gavage at a dose of 50mg/Kg, 100μL per dose, five-times per week, dissolved in ethanol, and delivered as a suspension in peanut oil, (c) rapamycin at a dose of 4mg/Kg 5-times a week by oral gavage, delivered as a suspension in peanut oil and (d) bicalutamide+rapamycin. At the end, mice were euthanized; and tumors collected for paraffin embedding. Blood from the euthanized animals was fractionated and serum collected for analysis.
D’Abronzo L.S., Bose S., Crapuchettes M.E., Beggs R.E., Vinall R.L., Tepper C.G., Siddiqui S., Mudryj M., Melgoza F.U., Durbin-Johnson B.P., deVere White R.W, & Ghosh. P.M. (2017). The Androgen Receptor is a negative regulator of eIF4E Phosphorylation at S209: Implications for the use of mTOR inhibitors in advanced prostate cancer. Oncogene, 36(46), 6359-6373.
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4

Prostate Tumor Xenograft Establishment and Analysis

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All experiments were conducted as approved by UC Davis IACUC Committee. 4–5-week old nu/nu athymic male mice were obtained from Harlan Sprague Dawley, Inc. (Indianapolis, IN) and implanted subcutaneously with sustained release testosterone pellets (12.5 mg, 90-day release; Innovative Research of America, Sarasota, FL). Suspensions of CWR22 or CWR22Rv1 cells were made in 50% Matrigel solubilized basement membrane (BD Biosciences, Bedford, MA) and xenografts were established by subcutaneous injections of 2.5 × 106 cells/site. When palpable tumors were observed, the testosterone pellets were removed and animals were followed for approximately four weeks, after which the mice were euthanized; and the tumors were collected. Part of the tumors were processed for paraffin embedding for immunohistochemistry, while the rest were lysed and homogenized for western blotting in cell lysis buffer (50 mM Tris HCl, pH 7.4, 150mM NaCl, and 1% NP-40, and protease inhibitors: 0.1mM benzamidine, 1mM phenylmethylsulfonyl fluoride, 10mg/ml each of phenathroline, leupeptin, aprotinin, and pepstatin A) and phosphatase inhibitors: 20mM β-glycerol phosphate, 1mM Na-orthovanadate, and 10mM NaF. Proteins were quantitated using a BCA assay (Pierce, Rockford IL) and fractionated on 29:1 acrylamide-bis SDS-PAGE.
Savoy R.M., Chen L., Siddiqui S., Melgoza F.U., Durbin-Johnson B., Drake C., Jathal M.K., Bose S., Steele T.M., Mooso B.A., D’Abronzo L.S., Fry W.H., Carraway KL I.I.I., Mudryj M, & Ghosh P.M. (2015). Transcription of Nrdp1 by the androgen receptor is regulated by nuclear Filamin A in prostate cancer. Endocrine-related cancer, 22(3), 369-386.
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5

Testosterone-Driven Xenograft Prostate Cancer Mouse Model

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4–5-week old nu/nu athymic male mice were obtained from Harlan Sprague Dawley, Inc. (Indianapolis, IN) and implanted subcutaneously with sustained release testosterone pellets (12.5 mg, 90-day release; Innovative Research of America). Suspensions of CWR22 cells were made in a 1:1 solution of RPMI 1640 (GIBCO) and Matrigel solubilized basement membrane (BD Biosciences). Xenografts were established by subcutaneous injections of 2.5 × 106 cells/injection into the right flank. When palpable tumors were observed, animals were (i) left intact (sham operated) or (ii) castrated by bilateral orchiectomy and removal of the testosterone pellet. The animals were followed for approximately four weeks, after which the mice were euthanized, and the tumors were resected. The study was conducted under an approved IACUC protocol.
Siddiqui S., Libertini S.J., Lucas C.A., Lombard A.P., Baek H.B., Nakagawa R.M., Nishida K.S., Steele T.M., Melgoza F.U., Borowsky A.D., Durbin-Johnson B.P., Qi L., Ghosh P.M, & Mudryj M. (2020). The p14ARF tumor suppressor restrains androgen receptor activity and prevents apoptosis in prostate cancer cells. Cancer letters, 483, 12-21.
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