Tour-de-Force

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Tour-de-Force: Interplay between Mitochondria and Cell Cycle Progression Fall 2007Appendix EE1 Cell Lines and cell culturesCell linesAll cell lines will be purchased through LGC Promochem in cooperation with ATCC unless statedotherwise.Mouse cell linesName ATCC # Cell type and tissue MotivationJ1 SCRC- Embryonic Stem Cell, Inner cell Control for proliferating cells1010 massMEF SCRC- Embryonic FibroblastDifferentiated stem cell(CF-1) 1040NIT-1 CRL- Insulinoma of the Islets of Might overexpress Cyclin D1 12055 LangerhansMOVAS CRL-2797Smooth Muscle of the Aorta Used in H1 and H3Human cell linesName ATCC # Cell type and tissue MotivationMaDo CCL-135 Lung fibroblast Non-proliferating cell(LL-47)HeLa CCL-2 Adenocarcinoma of the cervix Frequently used cell in researchA-704 HTB-45 Adenocarcinoma of the kidney Might overexpress cyclin D1 2WiDr CCL-218 Adenocarcinoma of the colon Might overexpresses cyclin D1 3Farage CRL- Non-Hodgkin B cell lymphoma Known to overexpress cyclin E 42630ZR-75-1 CRL- Ductal carcinoma (mammary Probably overexpress cyclin E 41500 gland)BC-3 CRL-2277B-cell lymphoma Known to overexpress cyclin D1 5The following cell line will be purchased through Promocell:HAoSMC-cC-12533 Human Aortic Smooth Muscle This is the cell line used in allhypothesisRat cell LinesName & ATCC # Cell type and tissue MotivationATCC #IA- CRL- Adenocarcinoma of the small To compare to HeLaXsSBR 1677 intestineRn1T Cancer of the mammary gland To compare to ZR-72-11 Chung et al., 20002 Schraml et al., 20033 Lin et al., 19984Tetsu & McCormick, 19995Motokura & Arnold, 1993SCI 332 Advanced Molecular Cell Biology Research Proposal 116
Tour-de-Force: Interplay between Mitochondria and Cell Cycle Progression Fall 2007Cell CulturesAll cell lines will be cultured at a temperature of 37º Celsius, the atmosphere will be 95% air and 5%carbon dioxide to maintain the pH. The medium and serum differs per cell line, the optimal growth mediaare listed below. The different components of the serum will be bought through ATCC unless indicatedotherwise. For subculturing protocols of all cell lines (except for HAoSMC-c) we would like to refer to theLGC website: www.lgcprochem-atcc.com. For HAoSMC-c cultering and thawing protocols we would like torefer to Promocell’s instructions ‘Recovery of crypopreserved cells’.J1 - This cell line will be propagated in Dulbecco’s modified Eagle’s Medium (DMEM) supplemented with2.0 mM L-Alanyl-L-Glutamine, 0.1 mM non-essential Amino Acids, 0.1 mM 2-mecrapoethanol (InvitrogenLife Technologies No. 21985), 1000 U/ml mouse leukemia inhibitory factor (LIF) (Chemicon No. ESG1107)and 15% fetal bovine serum.MEF - This cell line will be propagated in DMEM supplemented with with 4 mM L-glutamine adjusted tocontain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose (85%) and 15% fetal bovine serum.NIT-1 - This cell line will be propagated in Ham's F12K medium supplemented with 2 mM L-glutamineadjusted to contain 1.5 g/L sodium bicarbonate (90%) and 10% heat-inactivated dialyzed fetal bovineserum.MOVAS -This cell line will be propagated in DMEM with 4 mM L-glutamine adjusted to contain 1.5 g/Lsodium bicarbonate and 4.5 g/L glucose supplemented with 0.2 mg/ml G418, (90%) and 10% fetal bovineserum.MaDo - This cell line will be propagated in Ham’s F12K medium (85%) and 15% fetal bovine serum.HeLa, WiDr - These cell lines will be propagated in ATCC-formulated Eagle’s Minimum Essential Medium(EMEM) supplemented with fetal bovine serum to a final concentration of 10%.A-704 - This cell line will be propagated in EMEM in Earle’s BSS supplemented with nonessential aminoacids and sodium pyruvate (85%) and 15% fetal bovine serum.Farage, ZR-75-1 - These cell lines will be propagated in ATCC-formulated RPMI-1640 mediumsupplemented with fetal bovine serum to a final concentration of 10%.BC-3 - This cell line will be propagated in RPMI-1640 medium supplemented with 20% fetal bovine serum.HAoSMC-c – This cell line will be propagated in a vascular smooth muscle growth medium purchasedthrough Promocell will be used.IA-XsSBR - This cell line will be propagated in Ham’s F10 medium (90%) supplemented with 10% fetalbovine serum.Rn1T - This cell line will be propagated in DMEM supplemented with fetal bovine serum to a finalconcentration of 10%.SCI 332 Advanced Molecular Cell Biology Research Proposal 117
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