BE115+-+Lab02 - Lab 2: 2D and 3D Cell Culture 1...

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Page 1 of 6 Bioengineering 115: Cell Biology for Engineers Laboratory 2: 2D and 3D Cell Culture Last updated: January 21, 2011 Lab 2: 2D and 3D Cell Culture 1 Introduction In this laboratory you will subculture your cell culture from the last lab into a 2D and 3D culture preparation. The same will be done for a second cell line, which will be provided to you. The 3D culture will be prepared in agarose. In the next lab you will measure growth in each of the 4 subcultures and prepare to quantify proteins synthesized by the cells. You will note that this protocol is sparse compared the protocols for the last few labs. This is because this protocol draws heavily on skills you’ve learned in the past few labs. Active protocols in a research lab run the gamut between sparse and overflowing with detail. Often, a protocol will assume that you know how do something – in this lab, it is assumed that you know how to passage cells, so the full procedure is not described. This is one reason that it is critical to keep a detailed lab notebook. Before this lab, carefully review the sterile procedures from lab 1. 2 Objectives To prepare 2D and 3D cell cultures for later analysis. 3 References and Links NIH 3T3 Fibroblasts: Quantitative Studies of the Growth of Mouse Embryo Cells in Culture and Their Development Into Established Lines (http://jcb.rupress.org/content/17/2/299.abstract) Rex Chondrocytes: The Establishment and Characterization of an Immortal Cell Line With a Stable Chondrocytic Phenotype (http://onlinelibrary.wiley.com/doi/10.1002/jcb.10571/abstract) 4 Reagents, Supplies & Equipment 4.1 Reagents 1. 1x trypsin/EDTA solution (sterile) 0.05% trypsin 0.53 mM EDTA in Hanks Buffered Salt Solution 2. PBS (sterile) 3. 1.0% low melting point agarose in PBS (sterile) 4. Growth media - DMEM +10% calf serum, +1% pen/strep (sterile)
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Page 2 of 6 Bioengineering 115: Cell Biology for Engineers Laboratory 2: 2D and 3D Cell Culture Last updated: January 21, 2011 4.2 Supplies 1. T75 flask of fibroblasts from the last lab 2. Centrifuge tube of Rex cells 3. 50 mL sterile centrifuge tubes 4. 15 mL sterile centrifuge tubes 5. 1000 μL sterile pipet tips
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This note was uploaded on 10/14/2011 for the course BIOE 115 taught by Professor Haley during the Fall '11 term at City College of San Francisco.

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BE115+-+Lab02 - Lab 2: 2D and 3D Cell Culture 1...

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