Name: selective cell elimination from mixed 3d culture using a near infrared photoimmunotherapy technique
Uploaded: 2016-03-14T14:00:00
Description: Watch this Scientific Journal Video about Selective Cell Elimination from Mixed 3D Culture Using a Near Infrared Photoimmunotherapy Technique at JoVE.com

Denne forskningen ble støttet av egenutført Research Program av National Institutes of Health, National Cancer Institute, Senter for Cancer Research.

Merk: Følgende protokollen beskriver de nødvendige skritt for å eliminere bestemte celler ved hjelp av NIR-PIT. Kontroller og andre detaljer om NIR-PIT og celleviabilitet kan finnes andre steder 18. 1. Bøyning av IR700 til monoklonale antistoffer (mAb) <ol><li> Forbered mAb av interesse ved 2-5 mg / ml i 0,1 M Na HPO 2 4 (pH 8,6) oppløsning. </li><li> Bland 6,8 nmol av mAb med 30,8 nmol av 10 mM IR700 i 0,1 M Na HPO 2 4-løsning (pH 8,6) i et mikrosen...

<ol>
	<li>Robinton, D. A., Daley, G. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+promise+of+induced+pluripotent+stem+cells+in+research+and+therapy.">The promise of induced pluripotent stem cells in research and therapy.</a> Nature. 481 (7381), 295-305 (2012).</li><li>Yamanaka, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induced+pluripotent+stem+cells:+past,+present,+and+future.">Induced pluripotent stem cells: past, present, and future.</a> Cell stem cell. 10 (6), 678-684 (2012).</li><li>Birchall, M. A., Seifalian, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+engineering's+green+shoots+of+disruptive+innovation.">Tissue engineering's green shoots of disruptive innovation.</a> Lancet. 6736 (14), 11-12 (2014).</li><li>Ben-David, U., Benvenisty, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+tumorigenicity+of+human+embryonic+and+induced+pluripotent+stem+cells.">The tumorigenicity of human embryonic and induced pluripotent stem cells.</a> Nat. Rev. Cancer. 11 (4), 268-277 (2011).</li><li>Hanna, J. H., Saha, K., Jaenisch, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pluripotency+and+cellular+reprogramming:+facts,+hypotheses,+unresolved+issues.">Pluripotency and cellular reprogramming: facts, hypotheses, unresolved issues.</a> Cell. 143 (4), 508-525 (2010).</li><li>Lee, M. -. O., Moon, S. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+pluripotent+stem+cell-derived+teratoma+formation+by+small+molecules.">Inhibition of pluripotent stem cell-derived teratoma formation by small molecules.</a> Proc. Natl. Acad. Sci. U.S.A. 110 (35), 3281-3290 (2013).</li><li>Miura, K., Okada, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variation+in+the+safety+of+induced+pluripotent+stem+cell+lines.">Variation in the safety of induced pluripotent stem cell lines.</a> Nat. Biotechnol. 27 (8), 743-745 (2009).</li><li>Tang, C., Lee, A. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+antibody+against+SSEA-5+glycan+on+human+pluripotent+stem+cells+enables+removal+of+teratoma-forming+cells.">An antibody against SSEA-5 glycan on human pluripotent stem cells enables removal of teratoma-forming cells.</a> Nat. Biotechnol. 29 (9), 829-834 (2011).</li><li>Mitsunaga, M., Ogawa, M., Kosaka, N., Rosenblum, L. T., Choyke, P. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+cell+-+selective+in+vivo+near+infrared+photoimmunotherapy+targeting+specific+membrane+molecules.">Cancer cell - selective in vivo near infrared photoimmunotherapy targeting specific membrane molecules.</a> Nat. Med. 17 (12), 1685-1691 (2011).</li><li>Mitsunaga, M., Nakajima, T., Sano, K., Kramer-Marek, G., Choyke, P. L., Kobayashi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immediate+in+vivo+target-specific+cancer+cell+death+after+near+infrared+photoimmunotherapy.">Immediate in vivo target-specific cancer cell death after near infrared photoimmunotherapy.</a> BMC Cancer. 12 (1), 345 (2012).</li><li>Nakajima, T., Sano, K., Mitsunaga, M., Choyke, P. L., Kobayashi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Real-time+monitoring+of+in+vivo+acute+necrotic+cancer+cell+death+induced+by+near+infrared+photoimmunotherapy+using+fluorescence+lifetime+imaging.">Real-time monitoring of in vivo acute necrotic cancer cell death induced by near infrared photoimmunotherapy using fluorescence lifetime imaging.</a> Cancer Res. 72 (18), 4622-4628 (2012).</li><li>Sano, K., Mitsunaga, M., Nakajima, T., Choyke, P. L., Kobayashi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+cytotoxic+effects+of+photoimmunotherapy+assessed+by+18F-FDG+PET.">Acute cytotoxic effects of photoimmunotherapy assessed by 18F-FDG PET.</a> J. Nucl. Med. 54 (5), 770-775 (2013).</li><li>Sato, K., Watanabe, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photoimmunotherapy:+Comparative+effectiveness+of+two+monoclonal+antibodies+targeting+the+epidermal+growth+factor+receptor.">Photoimmunotherapy: Comparative effectiveness of two monoclonal antibodies targeting the epidermal growth factor receptor.</a> Mol. Oncol. 8 (3), 620-632 (2014).</li><li>Sato, K., Nagaya, T., Mitsunaga, M., Choyke, P. L., Kobayashi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near+infrared+photoimmunotherapy+for+lung+metastases.">Near infrared photoimmunotherapy for lung metastases.</a> Cancer Lett. 365 (1), 112-121 (2015).</li><li>Sato, K., Hanaoka, H., Watanabe, R., Nakajima, T., Choyke, P. L., Kobayashi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near+Infrared+Photoimmunotherapy+in+the+Treatment+of+Disseminated+Peritoneal+Ovarian+Cancer.">Near Infrared Photoimmunotherapy in the Treatment of Disseminated Peritoneal Ovarian Cancer.</a> Mol. Cancer Ther. 14 (8), 141-150 (2014).</li><li>Sato, K., Choyke, P. L., Kobayashi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photoimmunotherapy+of+Gastric+Cancer+Peritoneal+Carcinomatosis+in+a+Mouse+Model.">Photoimmunotherapy of Gastric Cancer Peritoneal Carcinomatosis in a Mouse Model.</a> PloS one. 9 (11), 113276 (2014).</li><li>Sato, K., Nagaya, T., Choyke, P. L., Kobayashi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near+Infrared+Photoimmunotherapy+in+the+Treatment+of+Pleural+Disseminated+NSCLC+Preclinical+Experience.">Near Infrared Photoimmunotherapy in the Treatment of Pleural Disseminated NSCLC Preclinical Experience.</a> Theranostics. 5 (7), 698-709 (2015).</li><li>Sato, K., Nakajima, T., Choyke, P. L., Kobayashi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+cell+elimination+in+vitro+and+in+vivo+from+tissues+and+tumors+using+antibodies+conjugated+with+a+near+infrared+phthalocyanine.">Selective cell elimination in vitro and in vivo from tissues and tumors using antibodies conjugated with a near infrared phthalocyanine.</a> RSC Adv. 5, 25105-25114 (2015).</li><li>Watanabe, R., Hanaoka, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photoimmunotherapy+Targeting+Prostate-Specific+Membrane+Antigen:+Are+Antibody+Fragments+as+Effective+as+Antibodies.">Photoimmunotherapy Targeting Prostate-Specific Membrane Antigen: Are Antibody Fragments as Effective as Antibodies.</a> J. Nucl. Med. 56 (1), 140-144 (2014).</li><li>Nakajima, T., Sano, K., Choyke, P. L., Kobayashi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improving+the+efficacy+of+Photoimmunotherapy+(PIT)+using+a+cocktail+of+antibody+conjugates+in+a+multiple+antigen+tumor+model.">Improving the efficacy of Photoimmunotherapy (PIT) using a cocktail of antibody conjugates in a multiple antigen tumor model.</a> Theranostics. 3 (6), 357-365 (2013).</li><li>Shirasu, N., Yamada, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potent+and+specific+antitumor+effect+of+CEA-targeted+photoimmunotherapy.">Potent and specific antitumor effect of CEA-targeted photoimmunotherapy.</a> Int J Cancer. 135 (11), 1-14 (2014).</li><li>Sato, K., Nagaya, T., Nakamura, Y., Harada, T., Choyke, P. L., Kobayashi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near+infrared+photoimmunotherapy+prevents+lung+cancer+metastases+in+a+murine+model.">Near infrared photoimmunotherapy prevents lung cancer metastases in a murine model.</a> Oncotarget. 6 (23), 19747-19758 (2015).</li><li>Nakajima, T., Sato, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+conjugate+and+light+dose+on+photo-immunotherapy+induced+cytotoxicity.">The effects of conjugate and light dose on photo-immunotherapy induced cytotoxicity.</a> BMC cancer. 14 (1), 389 (2014).</li><li>Klimanskaya, I., Rosenthal, N., Lanza, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Derive+and+conquer:+sourcing+and+differentiating+stem+cells+for+therapeutic+applications.">Derive and conquer: sourcing and differentiating stem cells for therapeutic applications.</a> Nat. Rev. Drug Discov. 7 (2), 131-142 (2008).</li><li>Burmester, G. R., Feist, E., D&#246;rner, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emerging+cell+and+cytokine+targets+in+rheumatoid+arthritis.">Emerging cell and cytokine targets in rheumatoid arthritis.</a> Nat. Rev. Rheumatol. 10 (2), 77-88 (2014).</li><li>Pardoll, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+blockade+of+immune+checkpoints+in+cancer+immunotherapy.">The blockade of immune checkpoints in cancer immunotherapy.</a> Nat. Rev. Cancer. 12 (4), 252-264 (2012).</li></ol>

Vi viser en metode for spesifikk celle eliminering fra en blandet 3D-cellekultur uten skade på ikke-målceller ved hjelp av NIR-PIT. Så langt er det ingen praktisk metode for eliminering celle når vev er etablert, eller etter transplantasjon. Således er NIR-PIT en lovende metode for å oppnå dette. Denne teknikken kan også bli anvendt in vivo 18,22, ettersom APCer viser lignende farmakokinetikk som mAb seg selv. Målcellen typen kan tilpasses med forskjellige APC. Forskjellige antistof...

Eliminere spesifikke celler uten å skade andre celler er ekstremt vanskelig, spesielt i etablert vev, og det er et stort behov for en celle eliminasjonsmetoden i tissue engineering feltet. Dag innen regenerativ medisin, vevkulturer ved hjelp av embryonale stamceller (ES), pluripotente stamceller (pscs), eller indusert pluripotent stamcelle (IPS) er lovende materialer 1 - 3. Selv om dette vev gjenfødelse er lovende, er forurensning med uønskede celler et stort problem. Videre er det et sikkerhetshensyn av tumorgenisitet etter transplantasjon 4,5. Selv om mange studier har fokusert på disse spørsmålene for å eliminere spesifikke celler, spesielt i regenerativ medisin 6-8, har ingen praktisk metode blitt utviklet. Nær infrarød photoimmunotherapy (NIR-PIT) er en behandling basert på et antistoff-photoabsorber konjugerte (APC). En APC består av en celle-spesifikt monoklonalt antistoff (mAb) og en photoabsorber, IR700. IR700 er en hydrofil silika-phthalocyanine derivat og ikke forårsaker fototoksisitet av seg selv 9. IR700 er kovalent konjugert til antistoffet via amid-rester på sidekjeden av lysin molekyler. APC binder målmolekylene på cellemembranen og induserer deretter nesten øyeblikkelig cellenekrose etter eksponering for NIR-lys ved 690 nm. Under eksponering for NIR-lys, cellemembranen brister som fører til celledød 9-14. NIR-PIT har vist seg å være effektiv med flere antistoffer eller antistoff-fragmenter, inkludert anti-EGFR, anti-HER2, anti-PSMA, anti-CD25, anti-mesothelin, anti-GPC3, og anti-CEA 15 - 21. Derfor kan NIR-PIT anvendes mot et bredt spekter av mål-molekyler. Videre er NIR-PIT en godt kontrollert behandling som gjør at selektiv behandling av spesifikke områder ved å begrense NIR-light bestråling 18,22. Her presenterer vi en metode for bestemt celle eliminering ved hjelp av NIR-PIT fra blandede 3D kulturer.

<table border="0" cellpadding="0" cellspacing="0" width="1083">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:424px;">IRDye 700DX Ester Infrared Dye</td>
			<td style="width:364px;">LI-COR Bioscience (Lincoln, NE, USA)</td>
			<td style="width:191px;">929-70011</td>
			<td style="width:104px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Na2HPO4</td>
			<td>SIGMA-ALDRICH (St. Louis, MO, USA)</td>
			<td>S9763</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sephadex G25 column (PD-10)&#160;</td>
			<td>GE Healthcare (Piscataway, NJ, USA)</td>
			<td>17-0851-01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Coomassie (bradford) Plus protein assay</td>
			<td>Thermo Fisher Scientific Inc (Waltham, MA, USA)</td>
			<td>PI-23200</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Perfecta3D 96-Well hanging Drop Plates</td>
			<td>3D Biomatrix Inc (Ann Arbor, MI, USA)</td>
			<td>HDP1096-8</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Optical power meter</td>
			<td>Thorlabs (Newton, NJ, USA)</td>
			<td>PM100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">LED: L690-66-60</td>
			<td>Marubeni America Co. (Santa Clara, CA, USA)</td>
			<td>L690-66-60</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vectibix (panitumumab)</td>
			<td>Amgen (Thousand Oaks, CA, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">35mm glass bottom dish, dish size 35mm, well size 10mm</td>
			<td>Cellvis (Mountain View, CA, USA)</td>
			<td>D35-10-0-N</td>
			<td></td>
		</tr>
	</tbody>
</table>

selective cell elimination from mixed 3d culture using a near infrared photoimmunotherapy technique

Recent developments in tissue engineering offer innovative solutions for many diseases. For example, tissue engineering using induced pluripotent stem cell (iPS) emerged as a new method in regenerative medicine. Although this tissue regeneration is promising, contamination with unwanted cells during tissue cultures is a major concern. Moreover, there is a safety concern regarding tumorigenicity after transplantation. Therefore, there is an urgent need for eliminating specific cells without damaging other cells that need to be protected, especially in established tissue. Here, we present a method for specific cell elimination from a mixed 3D cell culture in vitro with near infrared photoimmunotherapy (NIR-PIT) without damaging non-targeted cells. This technique enables the elimination of specific cells from mixed cell cultures or tissues.

Optisk å overvåke virkningen av NIR-PIT, A431-cellelinjen, som overuttrykker EGFR, var genetisk modifisert til å uttrykke også GFP og luciferase (A431-luc-GFP). Som et ikke-target av NIR-PIT, ble Balb / 3T3-cellelinje optisk modifisert for å uttrykke RFP (3T3-RFP). APC, panitumumab-IR700 (pan-IR700), ble syntetisert. Blandede sfæroider, som var sammensatt av forskjellige forhold mellom celler (A431-luc-GFP og 3T3-RFP) ble fremstilt i henhold til denne protokollen (figur 1)....

Watch this Scientific Journal Video about Selective Cell Elimination from Mixed 3D Culture Using a Near Infrared Photoimmunotherapy Technique at JoVE.com

Selective Cell Elimination from Mixed 3D Culture Using a Near Infrared Photoimmunotherapy Technique

Eliminating specific cells without damaging other cells is extremely difficult, especially in established tissue, yet there is an urgent need for a cell elimination method in the tissue engineering field. Here, we present a method for specific cell elimination from a mixed 3D cell culture using near infrared photoimmunotherapy (NIR-PIT).

Selektiv Cell Elimination fra Mixed 3D Kultur Ved hjelp av en nær-infrarødt Photoimmunotherapy Technique

Recent developments in tissue engineering offer innovative solutions for many diseases. For example, tissue engineering using induced pluripotent stem ...

The overall goal of this experimental procedure is to demonstrate the elimination of specific cells of interest from a mixed 3D cell culture using near infrared photoimmunotherapy. This method can help answer key questions in the tissue-engineering field about regenerating medicine. The main advantage of this technique that it can be used against a variety of target molecules and that the treatment can be restricted to specific lesions of interest.Thus can minimize the side effect. For IR700 conjugation to monoclonal antibodies, incubate the antibodies of interest with IR700 in Disodium Phosphate in a microcentrefuge tube at room temperature for one hour covered in aluminum foil. At the end of the incubation, wash a PD10 column two times with 15 milliliters of PBS.Then, load the antibody solution onto the column and purify the mixture by PBS elution according to the manufacturer's instructions. To confirm the number of fluoro-4 molecules conjugated to each monoclonal antibody molecule, use a spectrophotometer to determine the monoclonal antibody protein concentration. Then, store the conjugate at 4-degree Celsius.To prepare the mixed 3D cell culture, add 1 milliliter of sterile water into the plate reservoir section of a 96-well hanging drop plate. Then, plate various ratios of the cell types of interest in the appropriate cell culture medium at at least 5, 000 cells per 50 microliters of medium per well for five to seven days in a humidified incubator at 37-degree Celsius and 5%carbon dioxide. Change the culture medium every two days and observe the morphology and size of the spheroids daily with an inverted bright-field microscope at 10 to 40 times magnification.When the spheroids have reached 400 to 600 microns in diameter, replace the hanging drop plate medium with antibody photoabsorber conjugate-containing medium, and return the plate to the incubator for six hours. At the end of the incubation, wash the spheroids two times with fresh phenol red free-culture medium. Then use a 200-microliter pipette tip with a cut-off end to transfer one spheroid at a time into individual glass-bottom 50-millimeter dishes containing 100 microliters of fresh phenol red free-culture medium per dish.Use the inverted microscope to observe changes in the spheroid morphology. To irradiate the spheroids, place a light-emitting diode no higher than 5 inches above the glass-bottom dishes. And use an optical power meter to confirm the emission of a 670 to 710 nanometer wavelength at 2 joules per centimeter squared.After the NIR infrared photoimmunotherapy, transfer the spheroids into a new hanging drop plate with 50 microliters of fresh-culture medium per well for one day in the cell-culture incubator. Then, using another 200-microliter pipette tip with a cut-off end, gently transfer the spheroids into new glass-bottom dishes containing 100 microliters of phenol red free-culture medium, and assess the optical reporter expression under the flourescent microscope. Finally, detect the dead cells by the addition of propidium iodide to the medium.Or by the loss of cytoplasmic GFP flourescence. In this representative experiment, mixed spheroids composed of various ratios of A431-luc-GFP and 3T3-RFP cells were prepared as just demonstrated. Repeated NIR infrared photoimmunotherapy was then performed with the incubation of panitumumab-IR700 as illustrated.And the target cell elimination was monitored through the assessment of the flourescent and luciferase activities within the hanging drop cultures. Once mastered, this technique can be completed in one hour if it is performed properly. While attempting this procedure, it's important to remember that light energy decreases according to inverse-square law.

Research

JoVE Journal

Bioengineering

A Simple Hanging Drop Cell Culture Protocol for Generation of 3D Spheroids

Studies of cell-cell cohesion and cell-substratum adhesion have historically been performed on monolayer cultures adherent to rigid substrates. Cells ...

Rotating Cell Culture Systems for Human Cell Culture: Human Trophoblast Cells as a Model

Roterende Cell Kultur Systems for Human Cell Culture: Human trophoblast Cells som en modell

The field of human trophoblast research aids in  understanding the complex environment established during placentation. Due to the  nature of these ...

Preparation of 3D Fibrin Scaffolds for Stem Cell Culture Applications

Utarbeidelse av 3D Fibrin stillaser for Stem Cell Kultur Applications

Stem cells are found in naturally occurring 3D microenvironments in vivo, which are often referred to as the stem cell niche 1. Culturing stem cells ...

A Novel Method for Localizing Reporter Fluorescent Beads Near the Cell Culture Surface for Traction Force Microscopy

En ny metode for lokalisering Reporter fluoriserende perler Nær Cell Culture Surface for trekkraft Mikros

PA gels have long been used as a platform to study cell traction forces due to ease of fabrication and the ability to tune their elastic properties. When ...

Fluorescence-quenching of a Liposomal-encapsulated Near-infrared Fluorophore as a Tool for In Vivo Optical Imaging

Fluorescence-quenching of a Liposomal-encapsulated Near-infrared Fluorophore as a Tool for In Vivo Optical Imaging

Fluorescens-slukking av en Liposomalinnkapslet Nær-infrarød fluoroforen som et verktøy for<em&gt; I Vivo</em&gt; Optisk Imaging

Optical imaging offers a wide range of diagnostic modalities and has attracted a lot of interest as a tool for biomedical imaging. Despite the enormous ...

Tunable Hydrogels from Pulmonary Extracellular Matrix for 3D Cell Culture

Fleksibel Hydrogeler fra Lunge ekstracellulær matriks for 3D Cell Culture

Here we present a method for establishing multiple component cell culture hydrogels for in vitro lung cell culture. Beginning with healthy en bloc lung ...

Bioprinting of Cartilage and Skin Tissue Analogs Utilizing a Novel Passive Mixing Unit Technique for Bioink Precellularization

Bioprinting brusk og huden vev Analogs utnytte en roman passiv miksing enhet teknikk for Bioink Precellularization

Bioprinting is a powerful technique for the rapid and reproducible fabrication of constructs for tissue engineering applications. In this study, both ...

Fabrication of Custom Agarose Wells for Cell Seeding and Tissue Ring Self-assembly Using 3D-Printed Molds

Fabrikasjon av egendefinerte Agarose for celle såing og vev Ring selvstendig montering med 3D-trykt muggsopp

Engineered tissues are being used clinically for tissue repair and replacement, and are being developed as tools for drug screening and human disease ...

Construction of a Multilayered Mesenchymal Stem Cell Sheet with a 3D Dynamic Culture System

Byggingen av et flerlags Mesenchymal stamceller ark med en 3D dynamisk kultur-systemet

Stem cell therapy shows a promising future in regenerating injured organ and tissues, and the cell sheet technique has been developed to improve the low ...

Synthesis of Near-Infrared Emitting Gold Nanoclusters for Biological Applications

Syntese av nær-infrarød emitting gull nanoklynger for biologiske applikasjoner

Over the past decade, fluorescent gold nanoclusters (AuNCs) have witnessed growing popularity in biological applications and enormous efforts have been ...