Name: Silicon Microchips for Manipulating Cell-cell Interaction (Video) | JoVE Journal
Uploaded: 2007-08-30T00:00:00.000+00:00
Duration: 23 min 21 s
Description: Learn to manipulate cell-cell interactions using silicon micro mechanical chips with comb-like devices. This method enables dynamic regulation of hepatocyte and stromal cell communication for studying development and disease.

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The authors thank Salman Khetani, Jared Allen, Chris Flaim, and Austin Derfus for helpful discussions during the process of designing this device. This work was supported by the National Science Foundation Faculty Early Career Development Program, National Institutes of Health/National Institute of Diabetes and Digestive and Kidney Diseases, and the David and Lucile Packard Foundation. E.E.H. was supported by a Ruth L. Kirschstein National Research Service Award.

Preparation of cell cultures:
<ol>
<li>Start with silicon parts coated with plasma-treated polystyrene. </li>
<li>Coat parts with appropriate extracellular matrix proteins to support attachment of desired cell type. For hepatocytes, incubate in 50 g/ml Collagen-1 solution at 37&deg;C for 45 min. For 3T3 fibroblasts, no matrix is needed. </li>
<li>Lock parts with complementary parts in contact configuration. </li>
<li>Soak in 70% ethanol for a minimum of 10 min to sterilize. Rinse 2x in ddH2O, and 1x in cell culture media. </li>
<li>Seed cells at a concentration of 500,000 cells/ml in the appropriate culture medium. Use 1 ml in each well of a 12-well culture plate. </li>
<li>Incubate for 1 hr, shaking every 15 min to re-suspend cells evenly. </li>
<li>If a confluent monolayer has not been achieved after 1 hr, aspirate the cell suspension and repeat seeding with a fresh suspension. Repeat until the desired cell density has been achieved. </li>
<li>Remove the complementary parts. Transfer parts to fresh wells and incubate overnight to allow cells to adhere and spread fully. </li>
<li>Form co-cultures by locking appropriate parts into either the gap or contact configuration. The configuration may be changed at any desired point during culture. </li>
<li>When changing media, take care to leave cells dry for as little time as possible, preferably just a second or two. Draw out fluid with one hand and immediately replace media using the other hand. </li>
</ol>
Processing silicon parts for reuse:
<ol>
<li>Strip cells by soaking in bleach and rinsing with water. </li>
<li>Allow parts to dry completely. Soak in toluene for 2 h to strip polystyrene. </li>
<li>Clean in piranha solution (1:2 H2O2:H2SO4) heated to 120&deg;C. </li>
<li>Rinse for 15 min under a continuous flow of ddH2O. If you are not going to deposit polystyrene immediately, store the parts in water. </li>
<li>Dissolve polystyrene in toluene at 100 mg/ml. Vortex in polypropylene conical for about 1 hr, or until fully dissolved. A little more than 1 ml of solution is required for every 10 parts. </li>
<li>Spin coat polystyrene solution on individual silicon parts at 2,400 rpm for 30 sec. </li>
<li>Bake for at least 5 h at 120&deg;C. </li>
<li>Treat with oxygen plasma (200 mTorr, 200 W) for 1 min. </li>
</ol>

<ol><li>Hui, E. E., Bhatia, S. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Micromechanical+control+of+cell-cell+interactions%5BTitle%5D)+AND+%22Proceedings+of+the+National+Academy+of+Sciences%22%5BJournal%5D)">Micromechanical control of cell-cell interactions.</a> Proceedings of the National Academy of Sciences. 104, 
	
 <a target="_blank" href="http://www.ncbi.nlm.nih.gov/sites/entrez?Db=pubmed&Cmd=ShowDetailView&TermToSearch=17389399&ordinalpos=2&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum">http://www.ncbi.nlm.nih.gov/sites/entrez?Db=pubmed&Cmd=ShowDetailView&TermToSearch=17389399&ordinalpos=2&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum</a> 5722-5726 (2007).</li>
<li>Bhatia, S. N., Balis, U. J., Yarmush, M. L., Toner, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effect+of+cell-cell+interactions+in+preservation+of+cellular+phenotype%3A+cocultivation+of+hepatocytes+and+nonparenchymal+cells%5BTitle%5D)+AND+%22The+FASEB+Journal%22%5BJournal%5D)">Effect of cell-cell interactions in preservation of cellular phenotype: cocultivation of hepatocytes and nonparenchymal cells.</a> The FASEB Journal. 13, 
	
 <a target="_blank" href="http://www.ncbi.nlm.nih.gov/sites/entrez?Db=pubmed&Cmd=ShowDetailView&TermToSearch=10544172&ordinalpos=2&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum">http://www.ncbi.nlm.nih.gov/sites/entrez?Db=pubmed&Cmd=ShowDetailView&TermToSearch=10544172&ordinalpos=2&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum</a> 1883-1900 (1999).</li>
<li>El-Ali, J., Sorger, P. K., Jensen, K. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cells+on+Chips%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Cells on Chips.</a> Nature. 442, 
	
 <a target="_blank" href="http://www.ncbi.nlm.nih.gov/sites/entrez?Db=pubmed&Cmd=ShowDetailView&TermToSearch=16871208&ordinalpos=2&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum">http://www.ncbi.nlm.nih.gov/sites/entrez?Db=pubmed&Cmd=ShowDetailView&TermToSearch=16871208&ordinalpos=2&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum</a> 403-411 (2006).</li>
</ol>

The authors have nothing to disclose.

This system is unique in that it enables the spatial organization of tissue to be dynamically manipulated at the cellular level. Consequently, this device has enabled a number of novel biological experiments, spanning topics such as intercellular signaling dynamics, contact-mediated versus soluble signaling, cell fate decisions, toxicology, and cellular crosstalk. This device should be widely generalizable since the culture substrate is standard tissue culture plastic, and the system is compatible with standard culture m...

<table><tbody><tr><td>Silicon Comb Substrates</td><td>Tool</td><td>N/A</td><td>N/A</td><td>Parts are available for collaborative research projects. Please contact Elliot Hui (eehui @ alum . mit . edu) or Sangeeta Bhatia (sbhatia @ mit . edu).</td></tr></tbody></table>

silicon microchips for manipulating cell-cell interaction

Silicon Microchips for Manipulating Cell-cell Interaction

The role of the cellular microenvironment is recognized as crucial in determining cell fate and function in virtually all mammalian tissues from development to malignant transformation.&nbsp; In particular, interaction with neighboring stroma has been implicated in a plethora of biological phenomena; however, conventional techniques limit the ability to interrogate the spatial and dynamic elements of such interactions.
In Micromechanical Reconfigurable Culture (RC), we employ a micromachined silicon substrate with moving parts to dynamically control cell-cell interactions through mechanical repositioning. Previously, this method has been applied to investigate intercellular communication in co-cultures of hepatocytes and non-parenchymal cells, demonstrating time-dependent interactions and a limited range for soluble signaling 1.
Here, we describe in detail the preparation and use of the RC system. We begin by demonstrating the handling of the device parts using tweezers, including actuating between the gap and contact configurations (cell populations separated by a narrow 80-&micro;m gap, or in direct intimate contact). Next, we detail the process of preparing the substrates for culture, and the multi-step cell seeding process required for obtaining confluent cell monolayers. Using live microscopy, we then illustrate real-time manipulation of cells between the different possible experimental configurations. Finally, we demonstrate the steps required in order to regenerate the device surface for reuse: toluene and piranha cleaning, polystyrene coating, and oxygen plasma treatment.

This article describes an experimental approach for dynamic regulation of cell-cell interactions between adherent cells on a micrometer scale. Manipulation of intercellular communication between hepatocytes and stromal cell is demonstrated. The developed platform enables investigation of cell-cell interactions in a variety of biological processes, including development and pathogenesis.

The role of the cellular microenvironment is recognized as crucial in determining cell fate and function in virtually all mammalian tissues from ...

Hi, I am Elliot Hu.I'm a postdoctoral fellow in Tia's lab here at the Massachusetts Institute of Technology. Today we're gonna prepare cell cultures on silicon micro mechanical chips. The devices look like little combs that lock together and they allow us to precisely manipulate the interactions between two different populations of cells.Today we're gonna study the interaction between liver hepatocytes and supportive stromal cells. Specifically three T three fibroblasts cells are cultured on the top surface of the combs, so that by shifting the comb fingers, you can move the cells and change their spatial arrangement. Each device is divided into two parts which snap together in two possible configurations.The first brings the two cell populations into contact with each other. The second separates the two populations slightly so that the cells cannot touch. Here, contact interactions are prevented, but interactions through secreted Factors are preserved.Let's Start by talking About how to handle the parts with tweezers. I like to use both larger Teflon tweezers as well as smaller round tip metal tweezers. With the larger tweezers, you can pick up the parts on the edges like this wit

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Learn to manipulate cell-cell interactions using silicon micro mechanical chips with comb-like devices. This method enables dynamic regulation of hepatocyte and stromal cell communication for studying development and disease.

Silicon Microchips for Manipulating Cell-cell Interaction

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