Name: isolation of viable adipocytes and stromal vascular fraction from human visceral adipose tissue suitable for rna analysis and macrophage phenotyping
Uploaded: 2020-10-27T15:00:00
Description: Watch this Scientific Journal Video about Isolation of Viable Adipocytes and Stromal Vascular Fraction from Human Visceral Adipose Tissue Suitable for RNA Analysis and Macrophage Phenotyping at JoVE.c

Denne undersøgelse blev støttet af Instituto Nacional de Perinatologia (bevillingsnumre: 3300-11402-01-575-17 og 212250-3210-21002-06-15) og CONACyT, Fondo Sectorial de Investigacion en Salud y Seguridad Social (FOSISS) (bevillingsnummer 2015-3-2-61661).

Denne protokol blev godkendt af IRB ved Instituto Nacional de Perinatologia (212250-3210-21002-06-15). Deltagelse var frivillig, og alle de tilmeldte kvinder underskrev den informerede samtykkeformular.
1. Opsamling af visceralt fedtvæv
<ol><li>Få momsbiopsier gennem delvis omenektomi under kejsersnit fra raske voksne kvinder med singletongraviditeter på sigt uden arbejdskraft.</li>
	<li>Efter livmoderlukning og hæmostase, fortsæt med at identificere større omentum og udvide det på en...

Moms spiller en afgørende rolle i metabolisk regulering og inflammation. Stigende interesse for adipocytters og immuncellers rolle i kronisk inflammation forbundet med fedme har ført til udvikling af forskellige teknikker til at adskille SVF og fedtceller, der er til stede i AT. Imidlertid tillader de fleste teknikker ikke at opnå disse to forskellige sæt celler, der er levedygtige til downstream-applikationer fra den samme momsbiopsi i en enkelt procedure, hvilket kan være afgørende for undersøgelser vedrørende ...

Hvidt fedtvæv består ikke kun af fedtceller eller adipocytter, men også af en fedtfri cellefraktion kendt som stromal vaskulær fraktion (SVF), som indeholder en heterogen cellepopulation bestående af makrofager, andre immunceller som regulatoriske T-celler (Tregs) og eosinofiler, preadipocytter og fibroblaster, omgivet af vaskulært væv og bindevæv 1,2. Fedtvæv (AT) betragtes nu som et organ, der regulerer fysiologiske processer relateret til metabolisme og betændelse gennem adipokiner, cytokiner og mikroRNA'er produceret og frigivet af forskellige celler i vævet med autokrine, parakrin og endokrine virkninger 3,4. Hos mennesker omfatter hvidt fedtvæv det subkutane fedtvæv (SAT) og det viscerale fedtvæv (VAT) med vigtige anatomiske, molekylære, cellulære og fysiologiske forskelle mellem dem 2,5. SAT repræsenterer op til 80% af human AT, mens moms er placeret i bukhulen, hovedsageligt i mesenteriet og omentum, der metabolisk er mere aktiv6. Desuden er moms et endokrin organ, der udskiller mediatorer med en betydelig indvirkning på kropsvægt, insulinfølsomhed, lipidmetabolisme og inflammation. Derfor fører momsakkumulering til abdominal fedme og fedmerelaterede sygdomme såsom type 2-diabetes, metabolisk syndrom, hypertension og risiko for hjerte-kar-sygdomme, hvilket repræsenterer en bedre forudsigelse for fedmeassocieret dødelighed 6,7,8,9.
Under homeostatiske forhold samarbejder adipocytter, makrofager og de andre immunceller for at opretholde momsmetabolismen gennem udskillelsen af antiinflammatoriske mediatorer10. Imidlertid fremmer overdreven momsudvidelse rekrutteringen af aktiverede T-celler, NK-celler og makrofager. Faktisk er forholdet mellem makrofagerne i magert moms 5%, mens dette forhold stiger op til 50% i fedme, med makrofagpolarisering fra antiinflammatorisk til proinflammatorisk fænotype, hvilket genererer et kronisk inflammatorisk miljø10,11.
Som en konsekvens af fedmepandemien er der opstået et forbløffende antal rapporter, der behandler forskellige momsforskningsemner, herunder adipocytbiologi, epigenetik, inflammation, endokrine egenskaber og nye områder som ekstracellulære vesikler, blandt andet 8,10,12,13. Men selvom momsmiljøet er defineret ved krydstale mellem adipocytter og de residente eller ankommende makrofager, har de fleste undersøgelser kun fokuseret på en cellepopulation, og der er knappe oplysninger om interaktionen mellem disse celler i moms og deres patofysiologiske konsekvenser11,14. Desuden blev værdifulde undersøgelser, der adresserede samspillet mellem adipocyt og makrofag i AT, udført ved hjælp af cellelinjer, der manglede in vivo-primingbetingelserne 11,14,15. En passende strategi til at dissekere disse cellers interaktion eller særlige bidrag i moms kræver isolering af begge celletyper fra den samme fedtbiopsi for at udføre in vitro-assays, der afspejler så ens som muligt de in vivo-egenskaber, der regulerer momsmetabolismen.
Selvom de ikke-enzymatiske dissociationsmetoder baseret på mekaniske kræfter til at bryde AT sikrer minimal manipulation, kan disse metoder ikke bruges, hvis målet er at studere SVF-celler, da de har lavere effektivitet i cellegendannelse og lav cellelevedygtighed sammenlignet med enzymatiske metoder, og der er behov for et større volumen væv16,17. Enzymatisk fordøjelse ved hjælp af kollagenase er en skånsom metode, der tillader tilstrækkelig fordøjelse af kollagen og ekstracellulære matrixproteiner af fibrøst væv såsom WAT18 og bruges ofte, når trypsin er ineffektivt eller skadeligt19. Protokollen giver grundlæggende retningslinjer for fejlfinding for effektiv isolering af levedygtige modne adipocytter og SVF-celler fra humane momsbiopsier i en enkelt proces ved hjælp af en enzymatisk nedbrydningsteknik for kollagenase, der giver information for at sikre høje udbytter (mængde, renhed og integritet) af total RNA fra modne adipocytter, herunder mikroRNA'er, til downstream-ekspressionsapplikationer. Samtidig er protokollen optimeret til at identificere makrofagundergrupper fra SVF-celler gennem farvning af flere membranbundne markører til yderligere analyse ved flowcytometri20.

<table>
	<tbody>
		<tr>
			<td>0.2 mL PCR tubes</td>
			<td>Axygen</td>
			<td>PCR-02-C</td>
			<td>RNase, DNase free and nonpyrogenic</td>
		</tr>
		<tr>
			<td>1.5 mL microcentrifuge tubes</td>
			<td>Axygen</td>
			<td>MCT-150-C</td>
			<td>RNase, DNase free and nonpyrogenic</td>
		</tr>
		<tr>
			<td>10 mL serological pipettes</td>
			<td>Corning</td>
			<td>CLS4101-50EA</td>
			<td>Individually plastic wrapped</td>
		</tr>
		<tr>
			<td>10 &#181;L universal pipet tip</td>
			<td>Axygen</td>
			<td>T-300-L-R</td>
			<td>RNase, DNase free and nonpyrogenic</td>
		</tr>
		<tr>
			<td>10 &#181;L universal pipet tip</td>
			<td>Axygen</td>
			<td>T-300-R-S</td>
			<td>RNase, DNase free and nonpyrogenic</td>
		</tr>
		<tr>
			<td>1000 &#181;L universal pipet tip</td>
			<td>Axygen</td>
			<td>T-1000-B-R</td>
			<td>RNase, DNase free and nonpyrogenic</td>
		</tr>
		<tr>
			<td>2.0 mL microcentrifuge tube</td>
			<td>Axygen</td>
			<td>MCT-200-C</td>
			<td>RNase, DNase free and nonpyrogenic</td>
		</tr>
		<tr>
			<td>200 &#181;L universal pipet tip</td>
			<td>Axygen</td>
			<td>T-200-Y-R</td>
			<td>RNase, DNase free and nonpyrogenic</td>
		</tr>
		<tr>
			<td>2100 Bioanalyzer Instrument</td>
			<td>Agilent</td>
			<td>G2939BA</td>
			<td>-</td>
		</tr>
		<tr>
			<td>2101 Bioanalyzer PC</td>
			<td>Agilent</td>
			<td>G2953CA</td>
			<td>2100 Expert Software pre-installed in PC</td>
		</tr>
		<tr>
			<td>5 ml Round Bottom Polystyrene Test Tube</td>
			<td>Corning</td>
			<td>352003&#160;</td>
			<td>Snap cap, sterile</td>
		</tr>
		<tr>
			<td>50 mL centrifuge tubes</td>
			<td>Corning</td>
			<td>CLS430828-100EA</td>
			<td>Polipropilene, conical bottom and sterile</td>
		</tr>
		<tr>
			<td>Acid-guanidinium-phenol based reagent</td>
			<td>Zymo Research</td>
			<td>R2050-1-200</td>
			<td>TRI Reagent or similar</td>
		</tr>
		<tr>
			<td>Agilent RNA 6000 Nano Kit</td>
			<td>Agilent</td>
			<td>5067-1511</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Agilent Small RNA Kit</td>
			<td>Agilent</td>
			<td>5067-1548</td>
			<td>-</td>
		</tr>
		<tr>
			<td>APC/Cy7 anti-human CD14 Antibody</td>
			<td>BioLegend</td>
			<td>325620</td>
			<td>0.4 mg/106 cells, present on monocytes/macrophages, clone HCD14</td>
		</tr>
		<tr>
			<td>Baker</td>
			<td>-</td>
			<td>-</td>
			<td>250 ml, non sterile</td>
		</tr>
		<tr>
			<td>Bovine serum albumin</td>
			<td>Sigma-Aldrich</td>
			<td>A3912-100G</td>
			<td>Heat shock fraction, pH 5.2, &#8805;96%</td>
		</tr>
		<tr>
			<td>Chip priming station</td>
			<td>Agilent</td>
			<td>5065-9951</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Collagenase type II</td>
			<td>Gibco</td>
			<td>17101-015</td>
			<td>Powder</td>
		</tr>
		<tr>
			<td>D-(+)-Glucose</td>
			<td>Sigma-Aldrich</td>
			<td>G8270-100G</td>
			<td>Powder</td>
		</tr>
		<tr>
			<td>Direct-zol RNA Miniprep</td>
			<td>Zymo Research</td>
			<td>R2051</td>
			<td>Supplied with 50 mL TRI reagen</td>
		</tr>
		<tr>
			<td>Dissecting forceps</td>
			<td>-</td>
			<td>-</td>
			<td>Steel, serrated jaws and round ends</td>
		</tr>
		<tr>
			<td>Dissection tray</td>
			<td>-</td>
			<td>-</td>
			<td>Stainless steel</td>
		</tr>
		<tr>
			<td>Ethyl alcohol</td>
			<td>Sigma-Aldrich</td>
			<td>E7023-500ML</td>
			<td>200&#160;proof, for molecular biology</td>
		</tr>
		<tr>
			<td>FACS Flow Sheath Fluid</td>
			<td>BD Biosciences</td>
			<td>342003</td>
			<td>-</td>
		</tr>
		<tr>
			<td>FACS Lysing Solution</td>
			<td>BD Biosciences</td>
			<td>349202</td>
			<td>-</td>
		</tr>
		<tr>
			<td>FACSAria III Flow Cytometer/Cell Sorter</td>
			<td>BD Biosciences</td>
			<td>648282</td>
			<td>-</td>
		</tr>
		<tr>
			<td>FASCDiva Software</td>
			<td>BD Biosciences</td>
			<td>642868</td>
			<td>Software v6.0 pre-installed</td>
		</tr>
		<tr>
			<td>Hemacytometer</td>
			<td>Sigma</td>
			<td>Z359629-1EA</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Manual cell counter</td>
			<td>-</td>
			<td>-</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Mayo dissecting scisors</td>
			<td>-</td>
			<td>-</td>
			<td>Stainless steel</td>
		</tr>
		<tr>
			<td>Microcentrifuge</td>
			<td>-</td>
			<td>-</td>
			<td>Adjustable temperature</td>
		</tr>
		<tr>
			<td>Nanodrop spectrophotometer</td>
			<td>Thermo Scientific</td>
			<td>ND2000LAPTOP</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Orbital shaker</td>
			<td>-</td>
			<td>-</td>
			<td>Adjustable temperature and speed</td>
		</tr>
		<tr>
			<td>P10&#160;variable volume&#160;micropipette&#160;</td>
			<td>Thermo Scientific-Finnpipette</td>
			<td>4642040</td>
			<td>1 to 10 &#956;L</td>
		</tr>
		<tr>
			<td>P1000&#160;variable volume&#160;micropipette&#160;</td>
			<td>Thermo Scientific-Finnpipette</td>
			<td>4642090</td>
			<td>100 to 1000 &#956;L</td>
		</tr>
		<tr>
			<td>P2&#160;variable volume&#160;micropipette&#160;</td>
			<td>Thermo Scientific-Finnpipette</td>
			<td>4642010</td>
			<td>0.2 to 2 &#956;L</td>
		</tr>
		<tr>
			<td>P200&#160;variable volume&#160;micropipette&#160;</td>
			<td>Thermo Scientific-Finnpipette</td>
			<td>4642080</td>
			<td>20 to 200 &#956;L</td>
		</tr>
		<tr>
			<td>PCR tube storage rack</td>
			<td>Axygen</td>
			<td>R96PCRFSP</td>
			<td>-</td>
		</tr>
		<tr>
			<td>PE/Cy5 anti-human HLA-DR Antibody</td>
			<td>BioLegend</td>
			<td>307608</td>
			<td>0.0625 mg/106 cells, present on macrophages, clone L243</td>
		</tr>
		<tr>
			<td>PE/Cy7 anti-human CD45 Antibody</td>
			<td>BioLegend</td>
			<td>304016</td>
			<td>0.1 mg/106 cells, present on leukocytes, clone H130</td>
		</tr>
		<tr>
			<td>Phosphate buffered saline</td>
			<td>Sigma-Aldrich</td>
			<td>P3813-10PAK</td>
			<td>Powder, pH 7.4, for preparing 1 L solutions</td>
		</tr>
		<tr>
			<td>Pipette controller</td>
			<td>-</td>
			<td>-</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Red Blood Cells Lysis Buffer</td>
			<td>Roche</td>
			<td>11 814 389 001</td>
			<td>For preferential lysis of red blood cells from human whole blood</td>
		</tr>
		<tr>
			<td>Refrigerated centrifuge</td>
			<td>-</td>
			<td>-</td>
			<td>Whit adapter for 50 mL conical tubes</td>
		</tr>
		<tr>
			<td>Sterile Specimen container</td>
			<td>-</td>
			<td>-</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Transfer pipette</td>
			<td>Thermo Scientific-Samco</td>
			<td>204-1S</td>
			<td>Sterile</td>
		</tr>
		<tr>
			<td>Trypan Blue</td>
			<td>Gibco</td>
			<td>15250-061</td>
			<td>0.4% Solution</td>
		</tr>
		<tr>
			<td>Tube racks</td>
			<td>-</td>
			<td>-</td>
			<td>For different tube sizes</td>
		</tr>
		<tr>
			<td>Vortex Mini Shaker</td>
			<td>Cientifica SENNA</td>
			<td>BV101</td>
			<td>-</td>
		</tr>
	</tbody>
</table>

isolation of viable adipocytes and stromal vascular fraction from human visceral adipose tissue suitable for rna analysis and macrophage phenotyping

Visceralt fedtvæv (VAT) er et aktivt metabolisk organ, der hovedsageligt består af modne adipocytter og stromale vaskulære fraktionsceller (SVF), som frigiver forskellige bioaktive molekyler, der styrer metaboliske, hormonelle og immunprocesser; I øjeblikket er det uklart, hvordan disse processer reguleres i fedtvævet. Derfor er udviklingen af metoder, der evaluerer hver cellepopulations bidrag til fedtvævets patofysiologi, afgørende. Denne protokol beskriver isolationstrinnene og giver de nødvendige fejlfindingsretningslinjer for effektiv isolering af levedygtige modne adipocytter og SVF fra humane momsbiopsier i en enkelt proces ved hjælp af en enzymatisk nedbrydningsteknik for kollagenase. Desuden er protokollen også optimeret til at identificere makrofagundergrupper og udføre moden adipocyt-RNA-isolering til genekspressionsundersøgelser, hvilket gør det muligt at udføre undersøgelser, der dissekerer interaktionen mellem disse cellepopulationer. Kort fortalt vaskes momsbiopsier, hakkes mekanisk og fordøjes for at generere en enkeltcellesuspension. Efter centrifugering isoleres modne adipocytter ved flotation fra SVF-pelleten. RNA-ekstraktionsprotokollen sikrer et højt udbytte af total RNA (inklusive miRNA'er) fra adipocytter til downstream-ekspressionsassays. Samtidig bruges SVF-celler til at karakterisere makrofagundergrupper (pro- og antiinflammatorisk fænotype) gennem flowcytometrianalyse.

Denne protokol beskriver en enzymatisk metode, der anvender kollagenasefordøjelse efterfulgt af differentiel centrifugering til i en enkelt proces at isolere levedygtige modne adipocytter og SVF-celler fra momsbiopsier opnået fra raske gravide kvinder efter delvis omentektomi. I dette tilfælde bruger vi adipocytterne til RNA-ekstraktion og SVF til makrofagfænotypning.
RNA-ekstraktionsprotokollen gjorde det muligt at opnå RNA med en passende renhed, høj integritet og mikroRNA'er fra modne...

Watch this Scientific Journal Video about Isolation of Viable Adipocytes and Stromal Vascular Fraction from Human Visceral Adipose Tissue Suitable for RNA Analysis and Macrophage Phenotyping at JoVE.c

Isolation of Viable Adipocytes and Stromal Vascular Fraction from Human Visceral Adipose Tissue Suitable for RNA Analysis and Macrophage Phenotyping

Denne protokol tilvejebringer en effektiv kollagenasefordøjelsesmetode til isolering af levedygtige adipocytter og stromale vaskulære fraktion-SVF-celler fra humant visceralt fedt i en enkelt proces, herunder metode til opnåelse af RNA af høj kvalitet fra adipocytter og fænotypificering af SVF-makrofager gennem farvning af flere membranbundne markører til analyse ved flowcytometri.

Isolering af levedygtige adipocytter og stromal vaskulær fraktion fra humant visceralt fedtvæv, der er egnet til RNA-analyse og makrofagfænotypning

Visceral adipose tissue (VAT) is an active metabolic organ composed mainly of mature adipocytes and stromal vascular fraction (SVF) cells, which release ...

This protocol allows for isolation and study of viable adipocytes and stromal vascular fraction SVF cells from human visceral fat. Enzymatic digestion using collagenase ensures high yields of total RNA from mature adipocytes, including microRNAs suitable for expression analysis. Simultaneously, the method is optimized to identify macrophages by flow cytometry.This protocol provides fundamental troubleshooting guidelines for easy and efficient isolation of viable mature adipocytes and SVF cells from human VAT biopsies in a single process. After the uterine closure and hemostasis, identify greater omentum and extend it on a wet compress. Identify an avascular zone and use Kelly forceps to drill the outer side of visceral adipose tissue, or VAT.Use Ochsner forceps to clamp the proximal and distal side in the zone where VAT was drilled, and used Metzenbaum scissors to cut the tissue. Tie the greater omentum with black silk number one, then remove the Ochsner forceps and evaluate the hemostasis. To perform enzymatic digestion, cut four grams of VAT and use scissors to mince the tissue into small pieces in a dissection tray.Transfer the pieces to a new sterile 50-milliliter centrifuge tube and add 25 milliliters of digestion solution. Incubate the tube at 37 degrees Celsius for 60 minutes in an orbital shaker at 125 RPM. Then filter the digested tissue through three layers of gauze into a new sterile 50-milliliter centrifuge tube.Centrifuge the tube at 200 G for five minutes at four degrees Celsius. After centrifugation, two phases should be visible. The upper phase corresponds to mature adipocytes while SVF cells remain in the pellet.Transfer the mature adipocytes to a tube. Lyse cells by adding 1000 microliters of acid-guanidinium-phenol-based reagent and mix thoroughly with a P1000 micropipette to homogenize. Incubate the mixture for five minutes at room temperature to boost dissociation of nucleo-protein complexes.Centrifuge the cell lysate for five minutes at 12, 000 G to obtain three phases, an upper yellow phase corresponding to adipocyte lipids, a middle pink phase corresponding to nucleic acids, and a pellet with cellular debris. Carefully remove the lipid layer with a P200 micro pipette. Then transfer the middle phase into a new two-milliliter tube, taking care not to disturb the other phases.After purifying the RNA and measuring the concentration with a UV visible spectrophotometer, use an RNA integrity kit to determine the RNA integrity number. Then use the small RNA kit to determine the concentration and percent of microRNAs. Place the clip into a bioanalyzer.Dilute 10 microliters of SVF cell suspension into 90 microliters of 0.4%trypan blue solution, and apply 10 microliters to a standard hemocytometer. Count the viable cells carefully, excluding the dead cells, in four squares at the corner of the counting chamber. Transfer a total of 1 million SVF cells per milliliter to a five-milliliter round-bottom polypropylene test tube for flow cytometry and pellet the cells by centrifugation at 800 G for five minutes at four degrees Celsius.Vortex carefully to loosen the pellet and resuspend the cells in 100 microliters of PBS. Add the pre-titrated optimal concentration of each fluorochrome-conjugated monoclonal antibody specific for a cell surface antigen and mix gently, then incubate the cells for 15 minutes in the dark. Add an excess of cold PBS.And centrifuge the SVF at 400 G for five minutes at four degrees Celsius. Decant the supernatant, taking care to not disturb the pellet. Resuspend the pellet in five milliliters of sheath fluid.Then filter the cells through three layers of gauze into a new five-milliliter round-bottom polypropylene test tube. Vortex each tube briefly before analysis and count a minimum of 10, 000 events for flow analysis. This RNA extraction protocol was used to obtain adequate purity and high integrity RNA and microRNAs from mature adipocytes.The RNA integrity was assessed by an RNA quality control analyzer. Approximately 2.8 times 10 to the six nucleated cells were present per gram of visceral adipose tissue. The stromal vascular fraction cells were labeled with fluorophore-conjugated primary antibodies to identify and characterize 18 macrophages by fluorescence-activated cell-sorting analysis.Cell aggregates were eliminated from analysis by plotting the forward scatter area versus forward scatter height. Then cellular debris was excluded by gating the cells, based on the correct size and complexity, using forward scatter area and the side scatter area. To analyze macrophages, the monocyte and macrophage lineage cells were selected using CD45 and CD14 markers.Subsequently, the CD45-CD14 double positive cells were separated based on macrophage marker HLA-DR expression. Two subsets of macrophages were identified. In addition to identifying new cell phenotypes, this protocol makes it possible to use purified cell populations for future analyses, such as DNA methylation, post-translational modifications, and signaling pathways.

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Assessment of Human Adipose Tissue Microvascular Function Using Videomicroscopy

Vurdering af menneskers fedtvæv mikrovaskulære funktion ved hjælp af Videomicroscopy

While obesity is closely linked to the development of metabolic and cardiovascular disease, little is known about mechanisms that govern these processes. ...

Clinical Protocol of Producing Adipose Tissue-Derived Stromal Vascular Fraction for Potential Cartilage Regeneration

Klinisk protokol producere fedtvæv-afledte stromale vaskulære brøkdel for potentielle brusk regenerering

Osteoarthritis (OA) is one of the most common debilitating disorders.&#160;Recently, numerous attempts have been made to improve the functions of the ...

Isolation of Adipose Derived Regenerative Cells for the Treatment of Erectile Dysfunction Following Radical Prostatectomy

Isolering af fedtafledte regenerative celler til behandling af erektil dysfunktion efter radikal prostatektomi

Stem cells are used in many research areas within regenerative medicine in part because these treatments can be curative rather than symptomatic. Stem ...

Technique for Obtaining Mesenchymal Stem Cell from Adipose Tissue and Stromal Vascular Fraction Characterization in Long-Term Cryopreservation

Teknik til opnåelse af mesenkymal stamcelle fra fedtvæv og stromal vaskulær fraktionskarakterisering ved langvarig kryopræservering

Human mesenchymal stem cells derived from adipose tissue have become increasingly attractive as they show appropriate features and are an accessible ...

Isolation and Culture of Human Adipose-Derived Stem Cells With an Innovative Xenogeneic-Free Method for Human Therapy

Isolering og dyrkning af humane fedtafledte stamceller med en innovativ xenogenfri metode til human terapi

Considering the increasing impact of stem cell therapy, biosafety concerns have been raised regarding potential contamination or infection transmission ...