Name: antigens protected functional red blood cells by the membrane grafting of compact hyperbranched polyglycerols
Uploaded: 2013-01-02T12:00:00
Description: Watch this Scientific Journal Video about Antigens Protected Functional Red Blood Cells By The Membrane Grafting Of Compact Hyperbranched Polyglycerols at JoVE.com

Denne forskning er finansieret af de canadiske Blood Services (CBS) og den canadiske Institutes of Health Science (CIHR) Research Partnership Fund. Forfatterne takker LMB makromolekyle Hub på UBC Center for Blood forskning til brug af deres forskningsfaciliteter. Infrastrukturen facilitet er støttet af Canada fundament for Innovation (CFI) og Michael Smith Foundation for Health Research (MSFHR). R. Chapanian er modtager af (CIHR / CBS) ph.d.-stipendier inden for transfusionsmedicin Science og en modtager af MSFHR forskning praktikant post doc stipendium. JN Kizhakkedathu er modtager af MSFHR Career Investigator Scholar Award.

A. hyperforgrenede Polyglycerol Modification (SS-HPG) <ol><li> Sted lyofiliseret HPG 60 kDa (0,5 g, 0,0083 mmol) i en rundbundet kolbe og tørres natten over under vakuum ved 90 ° C. </li><li> Afkøles kolben til stuetemperatur, og opløs tørrede HPG i vandfrit pyridin (3 ml). </li><li> At funktionalisere cirka otte hydroxylgrupper på HPG med carboxylgrupper, tilsættes katalytisk mængde dimethylaminopyridin (en dråbe af 5 mg / ml opløsning i pyridin) til HPG opløsningen. Til denne blanding ravsyreanhydrid tils...

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Universal donor RBC har et stort potentiale i at øge blod tilgængelighed og sikkerhed for blodtransfusion terapi. RBC betragtes også lovende drug delivery køretøjer på grund af deres lange cirkulation og iboende biokompatibilitet 14, 15. Forsøg i dette papir evaluere in vitro-egenskaber HPG modificerede RBC&#39;er. Den in vitro-egenskaber og in vivo cirkulation af hpG modificerede RBCs er blevet undersøgt i vores gruppe for nylig 8, 11. Opdelingen af RBC&#39;er i ...

<table border="1"><tbody><tr><td> Glycidol </td><td> Sigma Aldrich </td><td></td><td> (ON, Canada) </td></tr><tr><td> Trimethylolpropane </td><td> Fluka </td><td></td><td> (ON, Canada) </td></tr><tr><td> Kaliummethylat </td><td> Sigma Aldrich </td><td></td><td> (ON, Canada) </td></tr><tr><td> Vandfrit pyridin </td><td> Sigma Aldrich </td><td></td><td> (ON, Canada) </td></tr><tr><td> 4-Dimethylaminopyridin </td><td> Sigma Aldrich </td><td></td><td> (ON, Canada) </td></tr><tr><td> Ravsyreanhydrid </td><td> Sigma Aldrich </td><td></td><td> (ON, Canada) </td></tr><tr><td> Acetone </td><td> Fisher Scientific </td><td></td><td> (ON, Canada) </td></tr><tr><td> Vandfrit dimethylformamid </td><td> Sigma Aldrich </td><td></td><td> (ON, Canada) </td></tr><tr><td> N-hydroxysuccinimid </td><td> Sigma Aldrich </td><td></td><td> (ON, Canada) </td></tr><tr><td> N, N&#39;-diisopropylcarbodiimid </td><td> Sigma Aldrich </td><td></td><td> (ON, Canada) </td></tr><tr><td> MTS-kort </td><td></td><td></td><td> Micro Typing System (MTS) kort (FL, USA) </td></tr><tr><td> Dextran 500 kDa </td><td> Pharmacia Fine Chemicals </td><td></td><td> (Sverige) </td></tr><tr><td> PEG 8 kDa </td><td> Sigma Aldrich </td><td></td><td> (ON, Canada) </td></tr><tr><td> FITC monoklonalt anti-Rhesus D (RhD) </td><td> Quotient Biodiagnostics </td><td></td><td> (PA, USA) </td></tr><tr><td> PE monoklonale anti-CD47 </td><td> BD Biosciences </td><td></td><td> (NJ, USA) </td></tr><tr><td> Drabkin reagens </td><td> Sigma Aldrich </td><td></td><td> (ON, Canada) </td></tr><tr><td></td><td></td><td></td><td> Tabel. Kemikalier og reagenser, der anvendes til podning af HPG polymerer til RBC-membran og deres analyse. </td></tr></tbody></table>

antigens protected functional red blood cells by the membrane grafting of compact hyperbranched polyglycerols

Røde blodlegemer (RBC) transfusion er afgørende for behandling af en række akutte og kroniske medicinske problemer såsom thalassæmi og seglcelle-anæmi 1-3. Som følge af tilstedeværelsen af mange antigener på RBC overflade (~ 308 kendte antigener 4), patienter i kronisk blodtransfusion terapi udvikler alloantistoffer grund af miss match af mindre antigener på transfunderet RBC&#39;er 4, 5. Podning af hydrofile polymerer, såsom polyethylenglycol (PEG) og hyperforgrenede polyglycerol (HPG) danner en udelukkelse lag på RBC membran, der forhindrer vekselvirkning af antistoffer med overfladeantigener uden at påvirke passagen af små molekyler såsom oxygen, glucose og ioner 3. I dag ingen metoder for frembringelsen af ​​universelle røde blodlegemer donorceller til dels på grund af den overvældende udfordring ved tilstedeværelsen af ​​store antal antigener (protein og kulhydrat baseret) på RBC overflade og ddvikling af sådanne metoder vil forbedre transfusion sikkerhed, og dramatisk forbedre adgangen til og brugen af ​​røde blodlegemer. I denne rapport er de eksperimenter, der bruges til at udvikle antigen beskyttede funktionelle RBC&#39;er af membranen podning af HPG og deres karakterisering præsenteret. HPGs er yderst biokompatible kompakte polymerer 6, 7, og forventes at være placeret i cellen glycocalyx, der omgiver lipidmembran 8, 9 og maske RBC overfladeantigener 10, 11.

Camouflere af Rhesus D antigen og CD47 RBC overfladeprotein blev kvantificeret ved strømningscytometri under anvendelse af fluorescerende mærkede monoklonale antistoffer, og et repræsentativt resultat er vist i figur 1.. Ved HPG-podede RBCs (grå), intensiteten af ​​signalet faldt (top forskudt mod venstre), sammenlignet med kontrollen RBC (røde og grønne), hvilket indikerer en reduktion i binding til antistoffer mod celleoverflade som angiver maskering af overfladeproteiner . <p class="jov...

Watch this Scientific Journal Video about Antigens Protected Functional Red Blood Cells By The Membrane Grafting Of Compact Hyperbranched Polyglycerols at JoVE.com

Antigens Protected Functional Red Blood Cells By The Membrane Grafting Of Compact Hyperbranched Polyglycerols

Cellemembranen modifikation af røde blodlegemer (RBC) med hyperbranched polyglycerol (HPG) præsenteres. Modificerede RBC blev karakteriseret ved vandig tofase partitionering, osmotisk skrøbelighed og komplement medieret lyse. Camouflage af overfladeproteiner og antigener blev evalueret under anvendelse af flowcytometri og Micro Typing System (MTS) blod fænotypebestemmelse kort.

Antigener Beskyttet Funktionelle røde blodlegemer ved The Membrane podning af Compact hyperforgrenet Polyglyceroler

Red blood cell (RBC) transfusion is vital for the treatment of a number of acute and chronic medical problems such as thalassemia major and sickle cell ...

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