Name: evaluating cell death using cell-free supernatant of probiotics in three-dimensional spheroid cultures of colorectal cancer cells
Uploaded: 2020-06-13T14:45:00
Description: Watch this Scientific Journal Video about Evaluating Cell Death Using Cell-Free Supernatant of Probiotics in Three-Dimensional Spheroid Cultures of Colorectal Cancer Cells at JoVE.com

Denna forskning stöddes av "Etablering av mätstandarder för kemi och strålning", bidragsnummer KRISS-2020-GP2020-0003 och "Utveckling av mätstandarder och teknik för biomaterial och medicinsk konvergens", bidragsnummer KRISS-2020-GP2020-0004-program, finansierade av Korea Research Institute of Standards and Science. Denna forskning stöddes också av ministeriet för vetenskap och IKT (MSIT), Koreas nationella forskningsstiftelse (NRF-2019M3A9F3065868), ministeriet för hälsa och välfärd (MOHW), Korea Health Industry Development Institute (KHIDI, HI20C0558), Handels-, industri- och energiministeriet (MOTIE) och Korea Evaluation Institute of Industrial Technology (KEIT, 20009350). ORCID ID (Hee Min Yoo: 0000-0002-5951-2137; Dukjin Kang: 0000-0002-5924-9674; Seil Kim: 0000-0003-3465-7118; Joo-Eun Lee: 0000-0002-2495-1439; Jina Lee: 0000-0002-3661-3701). Vi tackar Chang Woo Park för hjälp med experiment.

1. Bakteriella cellkulturer och beredning av Lactobacillus cellfri supernatant (LCFS)
OBS: Steg 1.2 – 1.9 utförs i en anaerob kammare.
<ol><li>Förbered en MRS agarplatta och buljong som innehåller L-cystein och sterilisera genom autoklavering.</li>
	<li>Förinkubera MRS-agarplattan i H2 anaerob kammare som hålls vid 37 °C med 20 ppm syre.</li>
	<li>Tina lactobacillus bakterielager och inokulera agarplattan med bakteriekulturen (Figur 1...

<ol>
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Vävnadsmikromiljön, inklusive närliggande celler och den extracellulära matrisen (ECM), är grundläggande för vävnadsgenerering och avgörande för kontrollen av celltillväxt och vävnadsutveckling13. 2D-kulturer har dock flera nackdelar, såsom störningar i cellinteraktioner, liksom förändringar i cellmorfologi, extracellulära miljöer och tillvägagångssättet i division14. 3D-cellkultursystem har studerats noggrant för att bättre reproducera in vivo-effekt...

Probiotika är de mest fördelaktiga mikroorganismerna i tarmen som förbättrar immunhomeostas och värdenergimetabolism1. Probiotika från Lactobacillus och Bifidobacterium är de mest avancerade i sitt slag som finns i tarmarna2,3. Tidigare undersökningar har visat att Lactobacillus har hämmande och antiproliferativa effekter på flera cancerformer, inklusive kolorektal cancer4. Dessutom förhindrar probiotika inflammatoriska tarmsjukdomar, Crohns sjukdom och ulcerös kolit5,6. De flesta studier med probiotika utfördes dock i tvådimensionella (2D) monoskikt som odlas på fasta ytor.
Artificiella odlingssystem saknar miljöegenskaper, vilket inte är naturligt för cancerceller. För att övervinna denna begränsning har tredimensionella (3D) odlingssystem utvecklats7,8. Cancerceller i 3D visar förbättringar när det gäller grundläggande biologiska mekanismer, såsom cell livskraft, spridning, morfologi, cellcellskommunikation, läkemedelskänslighet och in vivo relevans9,10. Dessutom är sfäroider gjorda av multicellulära typer av kolorektal cancer och är beroende av cellcellsinteraktioner och den extracellulära matrisen (ECM)11. Vår tidigare studie har rapporterat att probiotiska cellfria supernatant (CFS) som produceras med Lactobacillus fermentum visade anti-cancer effekter på 3D kulturer av kolorektal cancer (CRC) celler12. Vi föreslog att CFS är en lämplig alternativ strategi för att testa probiotiska effekter på 3D-sfäroider12.
Här presenterar vi ett tillvägagångssätt som kan rymma multicellulära typer av 3D-kolorektal cancer för analys av terapeutiska effekter av probiotiskt cellfritt supernatant (CFS) på flera 3D-kolorektal cancer mimicry system. Denna metod ger ett sätt att analysera relaterade probiotiska och anti-cancer effekter in vitro.

<table>
	<tbody>
		<tr>
			<td> 
			10% Mini-PROTEAN TGX Precast Protein Gels, 15-well, 15 &#181;l</td>
			<td>Biorad</td>
			<td>4561036</td>
			<td>Pkg of 10</td>
		</tr>
		<tr>
			<td>Applied Biosystems MicroAmp Optical Adhesive Film</td>
			<td>Thermo Fisher Scientific</td>
			<td>4311971</td>
			<td>100 covers</td>
		</tr>
		<tr>
			<td>10x transfer buffer</td>
			<td>Intron</td>
			<td>IBS-BT031A</td>
			<td>1 L</td>
		</tr>
		<tr>
			<td>10X Tris-Glycine (W/SDS)</td>
			<td>Intron</td>
			<td>IBS-BT014</td>
			<td>1 L</td>
		</tr>
		<tr>
			<td>Axygen 2.0 mL MaxyClear Snaplock Microcentrifuge Tube, Polypropylene, Clear, Nonsterile, 500 Tubes/Pack, 10 Packs/Case</td>
			<td>Corning</td>
			<td>SCT-200-C</td>
			<td>500 Tubes/Pack, 10 Packs/Case</td>
		</tr>
		<tr>
			<td>BD Difco Bacto Agar</td>
			<td>BD</td>
			<td>214010</td>
			<td>500 g</td>
		</tr>
		<tr>
			<td>BD Difco Lactobacilli MRS Broth</td>
			<td>BD</td>
			<td>DF0881-17-5</td>
			<td>500 g</td>
		</tr>
		<tr>
			<td>CellTiter-Glo 3D Cell viability assay</td>
			<td>Promega</td>
			<td>G9681</td>
			<td>100&#956;l/assay in 96-well plates</td>
		</tr>
		<tr>
			<td>Complete Protease Inhibitor Cocktail</td>
			<td>Sigma-Aldrich</td>
			<td>11697498001</td>
			<td>vial of 20 tablets</td>
		</tr>
		<tr>
			<td>Corning Phosphate-Buffered Saline, 1X without calcium and magnesium, PH 7.4 &#177; 0.1</td>
			<td>Corning</td>
			<td>21-040-CV</td>
			<td>500 mL</td>
		</tr>
		<tr>
			<td>EMD Millipore Immobilon-P PVDF Transfer Membranes</td>
			<td>fisher Scientific</td>
			<td>IPVH00010</td>
			<td>26.5cm x 3.75m roll; Pore Size: 0.45um</td>
		</tr>
		<tr>
			<td>Falcon 5 mL Round Bottom Polystyrene Test Tube, with Cell Strainer Snap Cap</td>
			<td>Corning</td>
			<td>352235</td>
			<td>25/Pack, 500/Case</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum, certified, US origin</td>
			<td>Thermo Fisher Scientific</td>
			<td>16000044</td>
			<td>500 mL</td>
		</tr>
		<tr>
			<td>iScript cDNA Synthesis Kit, 25 x 20 &#181;l rxns #1708890</td>
			<td>Biorad</td>
			<td>1708890</td>
			<td>25 x 20 &#181;L rxns</td>
		</tr>
		<tr>
			<td>iTaq Universal SYBR Green Supermix</td>
			<td>Biorad</td>
			<td>1725121</td>
			<td>5 x 1 mL</td>
		</tr>
		<tr>
			<td>Lactobacillus fermentum</td>
			<td>Korean Collection for Type Cultures</td>
			<td>KCTC 3112</td>
			<td></td>
		</tr>
		<tr>
			<td>L-Cysteine hydrochloride monohydrate</td>
			<td>Sigma-Aldrich</td>
			<td>C6852-25G</td>
			<td>25 g</td>
		</tr>
		<tr>
			<td>Methyl Cellulose (3500-5600mPa&#183;s, 2% in Water at 20&#176;C)</td>
			<td>TCI</td>
			<td>M0185</td>
			<td>500 g</td>
		</tr>
		<tr>
			<td>MicroAmp Fast Optical 96-Well Reaction Plate with Barcode, 0.1 mL</td>
			<td>Applied Biosystems</td>
			<td>4346906</td>
			<td>20 plates</td>
		</tr>
		<tr>
			<td>Millex-GS Syringe Filter Unit, 0.22 &#181;m, mixed cellulose esters, 33 mm, ethylene oxide sterilized</td>
			<td>Millipore</td>
			<td>SLGS033SB</td>
			<td>250</td>
		</tr>
		<tr>
			<td>PE Annexin V Apoptosis Detection Kit with 7-AAD</td>
			<td>Biolegend</td>
			<td>640934</td>
			<td>100 tests</td>
		</tr>
		<tr>
			<td>Penicillin-Streptomycin (10,000 U/mL)</td>
			<td>Thermo Fisher Scientific</td>
			<td>15140122</td>
			<td>100 mL</td>
		</tr>
		<tr>
			<td>Propidium Iodide</td>
			<td>Introgen</td>
			<td>P1304MP</td>
			<td>100 mg</td>
		</tr>
		<tr>
			<td>RIPA Lysis and Extraction Buffer</td>
			<td>Thermo Fisher Scientific</td>
			<td>89901</td>
			<td>250 mL</td>
		</tr>
		<tr>
			<td>RNeasy Mini Kit (250)</td>
			<td>Qiagen</td>
			<td>74106</td>
			<td>250</td>
		</tr>
		<tr>
			<td>RPMI-1640</td>
			<td>Gibco</td>
			<td>11875-119</td>
			<td>500 mL</td>
		</tr>
		<tr>
			<td>Trypsin-EDTA (0.25%), phenol red</td>
			<td>Thermo Fisher Scientific</td>
			<td>25200056</td>
			<td>100 mL</td>
		</tr>
		<tr>
			<td>Name of Materials/Equipment/Software</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments/Description</td>
		</tr>
		<tr>
			<td>anti - p-I&#954;B&#945; (B-9)</td>
			<td>Santa cruze</td>
			<td>sc-8404</td>
			<td>200 &#181;g/mL</td>
		</tr>
		<tr>
			<td>anti-BclxL (H-5)</td>
			<td>Santa cruze</td>
			<td>sc-8392</td>
			<td>200 &#181;g/mL</td>
		</tr>
		<tr>
			<td>anti-PARP 1 (C2-10)</td>
			<td>Santa cruze</td>
			<td>sc-53643</td>
			<td>50 &#181;l ascites</td>
		</tr>
		<tr>
			<td>anti-&#946;-actin (C4)</td>
			<td>Santa cruze</td>
			<td>sc-47778</td>
			<td>200 &#181;g/mL</td>
		</tr>
		<tr>
			<td>BD FACSVerse</td>
			<td>BD Biosciences</td>
			<td></td>
			<td>San Diego, CA, USA</td>
		</tr>
		<tr>
			<td>Synergy HTX Multi-Mode Microplate Reader</td>
			<td>BioT</td>
			<td>S1LFA</td>
			<td></td>
		</tr>
		<tr>
			<td>CO2 incubator</td>
			<td>Thermo fisher</td>
			<td>HERAcell 150i</td>
			<td></td>
		</tr>
		<tr>
			<td>Conical tube 15 ml</td>
			<td>SPL</td>
			<td>50015</td>
			<td></td>
		</tr>
		<tr>
			<td>Conical tube 50 ml</td>
			<td>SPL</td>
			<td>50050</td>
			<td></td>
		</tr>
		<tr>
			<td>Corning Costar Ultra-Low Attachment Multiple Well Plate</td>
			<td>Sigma-Aldrich</td>
			<td>CLS7007</td>
			<td></td>
		</tr>
		<tr>
			<td>Corning Costar Ultra-Low Attachment Multiple Well Plate</td>
			<td>Sigma-Aldrich</td>
			<td>CLS3471</td>
			<td></td>
		</tr>
		<tr>
			<td>Costar 50 mL Reagent Reservoirs, 5/Bag, Sterile</td>
			<td>Costar</td>
			<td>4870</td>
			<td></td>
		</tr>
		<tr>
			<td>Countess Cell Counting Chamber Slides</td>
			<td>Thermofisher</td>
			<td>C10228</td>
			<td></td>
		</tr>
		<tr>
			<td>Countess II FL Automated Cell Counter</td>
			<td>invitrogen</td>
			<td>AMQAF1000</td>
			<td></td>
		</tr>
		<tr>
			<td>EnSpire Multimode Reader</td>
			<td>Perkin Elmer</td>
			<td></td>
			<td>Enspire 2300</td>
		</tr>
		<tr>
			<td>Eppendorf Research Plus Multi Channel Pipette, 8-channel</td>
			<td>Eppendorf</td>
			<td>3122000051</td>
			<td></td>
		</tr>
		<tr>
			<td>FlowJo software</td>
			<td>TreeStar</td>
			<td></td>
			<td>Ashland, OR, USA</td>
		</tr>
		<tr>
			<td>Goat Anti-Mouse IgG (H+L)</td>
			<td>Jackson immunoresearch</td>
			<td>115-035-062</td>
			<td>1.5 mL</td>
		</tr>
		<tr>
			<td>Goat Anti-Rabbit IgG (H+L)</td>
			<td>Jackson immunoresearch</td>
			<td>111-035-144</td>
			<td>2.0 mL</td>
		</tr>
		<tr>
			<td>GraphPad Prism 5</td>
			<td>GraphPad Software</td>
			<td></td>
			<td>Inc., San Diego, CA, USA</td>
		</tr>
		<tr>
			<td>ImageJ</td>
			<td>NIH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>ImageQuant LAS 4000 mini</td>
			<td>Fujifilm</td>
			<td></td>
			<td>Tokyo, Japan</td>
		</tr>
		<tr>
			<td>Incubated shaker</td>
			<td>Lab companion</td>
			<td>SIF-6000R</td>
			<td></td>
		</tr>
		<tr>
			<td>Multi Gauge Ver. 3.0,</td>
			<td>Fujifilm</td>
			<td></td>
			<td>Tokyo, Japan</td>
		</tr>
		<tr>
			<td>Optical density (OD)LAMBDA UV/Vis Spectrophotometers</td>
			<td>Perkin Elmer</td>
			<td></td>
			<td>Waltham, MA, USA</td>
		</tr>
		<tr>
			<td>Phase-contrast microscope</td>
			<td>Olympus</td>
			<td></td>
			<td>Tokyo, Japan</td>
		</tr>
		<tr>
			<td>SPL microcentrifuge tube 1.5mL</td>
			<td>SPL</td>
			<td>60015</td>
			<td></td>
		</tr>
		<tr>
			<td>SPL Multi Channel Reservoirs, 12-Chs, PS, Sterile</td>
			<td>SPL</td>
			<td>21012</td>
			<td></td>
		</tr>
		<tr>
			<td>StepOnePlus Real-Time PCR system</td>
			<td>Thermo Fisher Scientific</td>
			<td></td>
			<td>Waltham, MA, USA</td>
		</tr>
		<tr>
			<td>Vibra-Cell Ultrasonic Liquid Processors</td>
			<td>SONICS-vibra cell</td>
			<td>VC 505</td>
			<td>500 Watt ultrasonic processor</td>
		</tr>
		<tr>
			<td>Vinyl Anaerobic Chamber</td>
			<td>COY LAB PRODUCTS</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

evaluating cell death using cell-free supernatant of probiotics in three-dimensional spheroid cultures of colorectal cancer cells

Detta manuskript beskriver ett protokoll för att utvärdera cancercellsdödsfall i tredimensionella (3D) sfäroider av multicellulära typer av cancerceller med hjälp av supernatanter från Lactobacillus fermentumcellkultur , som betraktas som probiotikakulturer. Användningen av 3D-kulturer för att testa Lactobacillus cellfria supernatant (LCFS) är ett bättre alternativ än att testa i 2D-monoskikt, särskilt som L. fermentum kan producera anti-cancereffekter i tarmen. L. fermentum supernatant identifierades att ha ökade anti-proliferative effekter mot flera kolorektal cancer (CRC) celler i 3D kultur villkor. Intressant nog var dessa effekter starkt relaterade till kulturmodellen, vilket visar den anmärkningsvärda förmågan hos L. fermentum att inducera cancercelldöd. Stabila sfäroider genererades från olika CRCs (kolorektal cancerceller) med hjälp av protokollet som presenteras nedan. Detta protokoll för att generera 3D-sfäroid är tidsbesparande och kostnadseffektivt. Detta system utvecklades för att enkelt undersöka anti-cancer effekter av LCFS i flera typer av CRC sfäroider. Som förväntat inducerade CRC sfäroider med LCFS starkt celldöd under experimentet och uttryckte specifika apoptos molekylära markörer som analyseras av qRT-PCR, western blotting och FACS analys. Därför är denna metod värdefull för att utforska cellens livskraft och utvärdera effekten av anti-cancerläkemedel.

Vi beskriver protokollet att erhålla sfäroider från olika kolorektal cancer cellinjer. Tillskott med metylcellulosa krävdes för att generera sfäroider. Vi presenterar också en metod för LCFS beredning och presentera en modell för att studera korrelationen mellan probiotika och kolorektal cancer. Sfäroidbildning och LCFS-förberedelseprotokoll illustreras schematiskt i figur 1A,B. Som visas i figur 2Aomvandlar metylcellulosakoncentratio...

Watch this Scientific Journal Video about Evaluating Cell Death Using Cell-Free Supernatant of Probiotics in Three-Dimensional Spheroid Cultures of Colorectal Cancer Cells at JoVE.com

Evaluating Cell Death Using Cell-Free Supernatant of Probiotics in Three-Dimensional Spheroid Cultures of Colorectal Cancer Cells

Här presenteras metoder för att förstå anti-cancer effekter av Lactobacillus cell-free supernatant (LCFS). Kolorektal cancer cellinjer visar celldöd när behandlas med LCFS i 3D kulturer. Processen att generera sfäroider kan optimeras beroende på byggnadsställningen och de analysmetoder som presenteras är användbara för att utvärdera de involverade signalvägarna.

Utvärdera celldöd med hjälp av cellfri supernatant av probiotika i tredimensionella sfäroidkulturer av kolorektalcancerceller

This manuscript describes a protocol to evaluate cancer cell deaths in three dimensional (3D) spheroids of multicellular types of cancer cells using ...

This protocol has been optimized to make it easier to investigate the anticancer effect of Lactobacillus cell-free supernatant in multiple types of colon cancer cell spheroids. This technique is the first approach that investigates the anti-cancer effects of LCFS on colon cancer spheroids under in vivo mimicry conditions. Begin by placing an autoclave sterilized MRS agar plate in a 37 degrees Celsius hydrogen anaerobic chamber with 20 parts per million oxygen and inoculating the plate with freshly thawed Lactobacillus bacterial stock.Add two to three milliliters of MRS broth, supplemented with l-cysteine to the tube and seal the tube with the butyl rubber stopper and tube cap. Next, use a loop to collect a single colony and place the colony into a 1.5 milliliter culture tube containing 500 microliters of PBS. Resuspend the colony homogenously within the saline and load the entire bacterial suspension into a one milliliter syringe.Insert the needle into the center of the culture tube lid and deliver the colony into the medium. Place the culture in a shaking incubator at 37 degrees Celsius, 5%carbon dioxide, and 200 revolutions per minute. After two days, measure the OD620 of the culture on a spectrophotometer.When the OD620 equals two, sediment the bacteria by centrifugation and wash the bacteria pellet with PBS. After the second centrifugation, resuspend the bacteria in four milliliters of RPMI 1640, supplemented with 10%fetal bovine serum without antibiotics and place the bacteria in the shaking incubator for four hours at 100 revolutions per minute. At the end of the incubation, pellet the bacteria by centrifugation and sterile filter the recovered supernatant through 0.22 micron strainer for minus 80 degrees Celsius storage.To prepare colorectal cancer cell lines for spheroid formation, culture the cells of interest as monolayers in 100 millimeter Petri dishes in the cell culture incubator. When the cells reach a 70 to 80%confluency, wash the cultures two times with four milliliters of PBS per wash before detaching the cells with one milliliter of 0.25%trypsin-EDTA per dish. After two minutes, check for dissociation under a light microscope.If the cells have detached, neutralize the enzymatic reaction with five milliliters of growth medium. Collect the cells by centrifugation in one 15 milliliter conical tube per cell culture and gently resuspend the pellets with three milliliters of growth medium per tube. Then, count the number of viable cells by trypan blue exclusion.After counting, dilute the cells to one to two times 10 to the fifth cells per milliliter concentration in fresh medium and add methylcellulose to the cells to a final 0.6%concentration. Transfer the cells to a sterile reservoir and use a multi-channel pipette to dispense 200 microliters of cells to each well of an ultra low attachment 96-well round bottom microplate. Then, place the plate in the cell culture incubator for 24 to 36 hours before checking for the formation of spheroids by light microscopy.For LCFS treatment, serially dilute the prepared thawed LCFS stock solution in fresh growth medium to 25, 12.5, and 6%concentrations and use a 200 microliter pipette to carefully remove as much supernatant as possible from each well of the plated colorectal cancer cell line cultures. Then, gently add 200 microliters of LCFS supplemented growth medium in triplicate to each set of cancer cells and return the plate to the incubator for an additional 24 to 48 hours. A methylcellulose concentration of 0.6%transforms the tested cancer cell line cells into compact spheroids, indicating that spheroids can be generated from several types of colorectal cancer using this protocol.Spheroids cultured with 25%LCFS exhibit disrupted surfaces and a decreased cell viability in a dose-dependent manner. Indeed, LCFS co-cultured cells demonstrate higher levels of propidium iodide expression compared to viable control cells. Reverse transcriptase PCR analysis, Western blotting and flow cytometric annexin V/7-AAD analysis can also be performed to assess the effects of LCFS on cancer cell apoptosis.Be sure to handle the spheroids gently when removing the growth medium as the spheroids can easily break, making it impossible to observe the effect of LCFS treatment. Following this procedure, cell viability assays, quantitative real-time PCR, Western blotting and FACS analysis can be performed to characterize the anticancer effect of LCFS in greater detail.

Research

JoVE Journal

Cancer Research

Utilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Cultures

Med hjälp av funktionsgenomik screening för att identifiera potentiellt ny läkemedelsmål för cancercellernas Spheroid Kulturer

The identification of functional driver events in cancer is central to furthering our understanding of cancer biology and indispensable for the discovery ...

Preparation and Metabolic Assay of 3-dimensional Spheroid Co-cultures of Pancreatic Cancer Cells and Fibroblasts

Förberedelse och metabola haltbestämning av 3-dimensionell sfäroid samtidig kulturer av pankreascancer celler och fibroblaster

Many cancer types, including pancreatic cancer, have a dense fibrotic stroma that plays an important role in tumor progression and invasion. Activated ...

Moving Upwards: A Simple and Flexible In Vitro Three-dimensional Invasion Assay Protocol

Moving Upwards: A Simple and Flexible In Vitro Three-dimensional Invasion Assay Protocol

Flytta uppåt: Ett enkelt och flexibelt In Vitro tredimensionella Invasion Assay protokoll

Although 3D invasion assays have been developed, the challenge remains to study cells without affecting the integrity of their microenvironment. ...

Fully Human Tumor-based Matrix in Three-dimensional Spheroid Invasion Assay

Helt mänsklig tumör-baserade Matrix i tredimensionell sfäroid invasion assay

Two-dimensional cell culture-based assays are commonly used in in vitro cancer research. However, they lack several basic elements that form the tumor ...

Assessing Cell Viability and Death in 3D Spheroid Cultures of Cancer Cells

Bedömning cell livs kraft och död i 3D Sfäroiden kulturer av cancer celler

Three-dimensional spheroids of cancer cells are important tools for both cancer drug screens and for gaining mechanistic insight into cancer cell biology. ...

Evaluating the Differentiation Capacity of Mouse Prostate Epithelial Cells Using Organoid Culture

Utvärdering av differentierings kapacitet av mus prostata epitelceller med Organoid kultur

The prostate epithelium is comprised predominantly of basal and luminal cells. In vivo lineage tracing has been utilized to define the differentiation ...

Isolation of Stem-like Cells from 3-Dimensional Spheroid Cultures

Isolering av Stem-liknande celler från 3-dimensionella sfäroid kulturer

Despite advances in adult stem cell research, identification and isolation of stem cells from tissue specimens remains a major challenge. While resident ...

Detection of Cell-Free DNA in Blood Plasma Samples of Cancer Patients

Påvisande av cellfritt DNA i blodplasmaprover från cancerpatienter

Identifying mutations in tumors of cancer patients is a very important step in disease management. These mutations serve as biomarkers for tumor diagnosis ...

A Three-dimensional Model of Spheroids to Study Colon Cancer Stem Cells

En tredimensionell modell av sfäroider för att studera tjocktarmscancerstamceller

Colorectal cancers are characterized by heterogeneity and a hierarchical organization comprising a population of cancer stem cells (CSCs) responsible for ...

Three-Dimensional In Vitro Biomimetic Model of Neuroblastoma Using Collagen-Based Scaffolds

Three-Dimensional In Vitro Biomimetic Model of Neuroblastoma Using Collagen-Based Scaffolds

Tredimensionell in vitro biomimetisk modell av neuroblastom med hjälp av kollagenbaserade strukturer

Neuroblastoma is the most common extracranial solid tumor in children, accounting for 15% of overall pediatric cancer deaths. The native tumor tissue is a ...