Name: rapid isolation of bmpr-ib+ adipose-derived stromal cells for use in a calvarial defect healing model
Uploaded: 2017-02-24T13:00:00
Description: Watch this Scientific Journal Video about Rapid Isolation of BMPR-IB+ Adipose-Derived Stromal Cells for Use in a Calvarial Defect Healing Model at JoVE.com

C.D.M. was supported by the American College of Surgeons (ACS) Resident Research Scholarship. M.S.H. was supported by the California Institute for Regenerative Medicine (CIRM) Clinical Fellow training grant TG2-01159. M.S.H., H.P.L., and M.T.L. were supported by the American Society of Maxillofacial Surgeons (ASMS)/Maxillofacial Surgeons Foundation (MSF) Research Grant Award. H.P.L. was supported by NIH grant R01 GM087609 and a gift from Ingrid Lai and Bill Shu in honor of Anthony Shu. H.P.L. and M.T.L. were supported by the Hagey Laboratory for Pediatric Regenerative Medicine and The Oak Foundation. M.T.L. was supported by NIH grants U01 HL099776, R01 DE021683-01, and RC2 DE020771. D.C.W. was supported by NIH grant 1K08DE024269, the Hagey Laboratory for Pediatric Regenerative Medicine, and the Stanford University Child Health Research Institute Faculty Scholar Award.

Not: numuneleri rıza veren hastadan elde edildi. Tüm protokoller gözden geçirilmiş ve uygun Stanford Üniversitesi Kurumsal Değerlendirme Kurulu tarafından onaylanmıştır. insan doku ve hücrelerinin işlerken, her zaman kurum tarafından belirtilen, Biyogüvenlik Seviyesi 2 (BSL2) önlemlere uymak. Reaktiflerin 1. Hazırlık <ol><li> FACS tampon hazırlanması: 10 ml FBS, 188 5 ml Poloxamer 500 ml steril fosfat tamponlu tuz için 5 ml Pen-Strep (PBS) ekleyin. </li><li> Karışım...

<ol>
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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adipose-derived+stem+cells+for+regenerative+medicine.">Adipose-derived stem cells for regenerative medicine.</a> Circ Res. 100 (9), 1249-1260 (2007).</li><li>Levi, B., 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=CD105+protein+depletion+enhances+human+adipose-derived+stromal+cell+osteogenesis+through+reduction+of+transforming+growth+factor+beta1+(TGF-beta1)+signaling.">CD105 protein depletion enhances human adipose-derived stromal cell osteogenesis through reduction of transforming growth factor beta1 (TGF-beta1) signaling.</a> J Biol Chem. 286 (45), 39497-39509 (2011).</li><li>Chung, M. 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C., 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=Osteogenic+differentiation+of+mouse+adipose-derived+adult+stromal+cells+requires+retinoic+acid+and+bone+morphogenetic+protein+receptor+type+IB+signaling.">Osteogenic differentiation of mouse adipose-derived adult stromal cells requires retinoic acid and bone morphogenetic protein receptor type IB signaling.</a> Proc Natl Acad Sci U S A. 103 (33), 12335-12340 (2006).</li><li>McArdle, A., 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=Positive+selection+for+bone+morphogenetic+protein+receptor+type-IB+promotes+differentiation+and+specification+of+human+adipose-derived+stromal+cells+toward+an+osteogenic+lineage.">Positive selection for bone morphogenetic protein receptor type-IB promotes differentiation and specification of human adipose-derived stromal cells toward an osteogenic lineage.</a> Tissue Eng Part A. 20 (21-22), 3031-3040 (2014).</li><li>Sharon, Y., Alon, L., Glanz, S., Servais, C., Erez, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+normal+and+cancer-associated+fibroblasts+from+fresh+tissues+by+Fluorescence+Activated+Cell+Sorting+(FACS).">Isolation of normal and cancer-associated fibroblasts from fresh tissues by Fluorescence Activated Cell Sorting (FACS).</a> J Vis Exp. 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Protokol çerçevesinde kritik adımlar TSK Hasat sırasında, kritik bir adım kollajenaz ile yağ yeterli sindirim olduğunu. Yetersiz sindirim TSK düşük bir verimle sonuçlanır. BMPR-IB + hücrelerinin FACS sıralama sırasında, dikkatli pozitifliği açısından kapısı tanımlamak önemlidir. çok gevşek kapıları tanımlanması saf değildir sıralı nüfus neden olabilir. kalvarial defekt oluşturulması sırasında, kafatası kemik yoluyla kusur delmek için ancak dura mater iç...

yaralanma, enfeksiyon veya invaziv kanser kaynaklanan başlıca kemik defektleri hastanın iyileşmesi ve yaşam kalitesi üzerinde önemli bir etkiye sahip. Teknikleri yerde hastanın kendi vücudunda sağlıklı kemik ile bu kusurları doldurmak için var, ama bu transfer, kendi morbidite ve komplikasyonları 1, 2, 3 riskini taşır. Ayrıca, bazı kusurları yeterli donör kemik defekti doldurmak için mevcut değildir o kadar büyük veya karmaşık. Protez cihazlar kemikli defektleri doldurmak için potansiyel bir seçenek ancak bu enfeksiyon riski, donanım arızası ve yabancı cisim reaksiyonu 4 dahil olmak üzere çeşitli dezavantajları ile ilişkilidir. Bu nedenlerden dolayı hastanın kendi hücrelerini 5 kullanan biyolojik kemik yerine geçen mühendislik olasılığı büyük ilgi var. Yağ-kaynaklı stromal hücreler (TSK)onlar hastanın kendi yağ dokusunda bolca mevcuttur ve yeni kemik dokusu 6, 7 üreterek kemik defektleri iyileşmek için yeteneğini göstermiştir çünkü bu uygulama için potansiyel var. TSK hücreler ve çeşitli çalışmalar çeşitli nüfus spesifik hücre yüzey belirteçleri için seçilerek geliştirilmiş osteojenik aktivitesi 8, 9 hücre popülasyonları göstermiştir vardır. En yüksek osteojenik potansiyele sahip TSK seçilmesi, bu hücreleri ile seribaşı bir iskele büyük bir kemik defekti yeniden olabilir olasılığını artırmak olacaktır. Kemik morfogenetik proteini (BMP) sinyal kemik farklılaşmasını ve oluşumu 10 ve TSK 11 osteogenesis için önemli olduğu bilinen BMP reseptörü tip lb (BMPR-IB) düzenlenmesi için çok önemlidir. Son zamanlarda, biz olabilir b BMPR-IB o ifadeyi göstermiştirE geliştirilmiş osteojenik aktivite 12 TSK için seçmek için kullanılır. Burada, bir in vivo kalvaryal hata modeli kullanarak osteojenik aktivitesi deneyi takiben, insan yağ TSK BMPR-IB-sentezleyen izolasyonu için protokol göstermektedir.

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rapid isolation of bmpr-ib+ adipose-derived stromal cells for use in a calvarial defect healing model

Invasive cancers, major injuries, and infection can cause bone defects that are too large to be reconstructed with preexisting bone from the patient's own body. The ability to grow bone de novo using a patient's own cells would allow bony defects to be filled with adequate tissue without the morbidity of harvesting native bone. There is interest in the use of adipose-derived stromal cells (ASCs) as a source for tissue engineering because these are obtained from an abundant source: the patient's own adipose tissue. However, ASCs are a heterogeneous population and some subpopulations may be more effective in this application than others. Isolation of the most osteogenic population of ASCs could improve the efficiency and effectiveness of a bone engineering process. In this protocol, ASCs are obtained from subcutaneous fat tissue from a human donor. The subpopulation of ASCs expressing the marker BMPR-IB is isolated using FACS. These cells are then applied to an in vivo calvarial defect healing assay and are found to have improved osteogenic regenerative potential compared with unsorted cells.

Mikro BT açıkça kafatası defekti gösterecektir ameliyat günü yapılan tarama. Şu anda 4 mm defekt içine hiçbir büyümesi olacak. Daha sonraki taramaları başlangıca kıyasla zamanla kusurun büyüklüğünü ölçmek için zamanla elde edilir. BMPR-IB- ve tasnif hücreleri (Şekil 5) ile karşılaştırıldığında BMPR IB + hücreleri ile tohumlanmış Kusur kusurun daha hızlı kapanmasına göstermelidir. Buna ek olarak, kusur içeren kafatasının kısm...

Watch this Scientific Journal Video about Rapid Isolation of BMPR-IB+ Adipose-Derived Stromal Cells for Use in a Calvarial Defect Healing Model at JoVE.com

Rapid Isolation of BMPR-IB+ Adipose-Derived Stromal Cells for Use in a Calvarial Defect Healing Model

Adipose-derived stromal cells may be useful for engineering new tissue from a patient's own cells. We present a protocol for the isolation of a subpopulation of human adipose-derived stromal cells (ASCs) with increased osteogenic potential, followed by application of the cells in an in vivo calvarial healing assay.

BMPR-IB + hızlı İzolasyon Kalvaryum Hata Şifa Modeli Kullanılan Stromal Hücreleri yağ kaynaklı

Invasive cancers, major injuries, and infection can cause bone defects that are too large to be reconstructed with preexisting bone from the patient's own ...

Research

JoVE Journal

Developmental Biology

Isolation and Enrichment of Human Adipose-derived Stromal Cells for Enhanced Osteogenesis

İzolasyon ve İnsan zenginleştirilmesi Gelişmiş Osteogenezi için Stromal Hücreleri adipoz-türevi

Bone marrow-derived mesenchymal stromal cells (BM-MSCs) are considered the gold standard for stem cell-based tissue engineering applications. However, the ...

Automated Quantification of Hematopoietic Cell &#8211; Stromal Cell Interactions in Histological Images of Undecalcified Bone

Automated Quantification of Hematopoietic Cell ÃƒÂ¢Ã¢â€šÂ¬Ã¢â‚¬Å“ Stromal Cell Interactions in Histological Images of Undecalcified Bone

Hematopoetik Hücre Otomatik Ölçümü - undekalsifiye Kemik histolojik Görüntüler stromal hücre etkileşimleri

Confocal microscopy is the method of choice for the analysis of localization of multiple cell types within complex tissues such as the bone marrow. ...

Isolation and Differentiation of Adipose-Derived Stem Cells from Porcine Subcutaneous Adipose Tissues

Domuz Subkutan yağ dokularındaki İzolasyonu ve Yağ-Türetilmiş Kök Hücre Farklılaşması

Obesity is an unconstrained worldwide epidemic. Unraveling molecular controls in adipose tissue development holds promise to treat obesity or diabetes. ...

Immunomagnetic Separation of Fat Depot-specific Sca1high Adipose-derived Stem Cells (ASCs)

Immunomagnetic Separation of Fat Depot-specific Sca1high Adipose-derived Stem Cells ASCs

Yağ Deposu özgü SCA1 ve İmmunomanyetik Ayırma<sup&gt; yüksek</sup&gt; Yağ türetilmiş Kök Hücreler (TSK)

The isolation of adipose-derived stem cells (ASCs) is an important method in the field of adipose tissue biology, adipogenesis, and extracellular matrix ...

Isolation and Expansion of Mesenchymal Stem/Stromal Cells Derived from Human Placenta Tissue

İnsan Plasenta Doku türetilen İzolasyon ve mezenkimal kök Genişleme / Stromal hücreler

Mesenchymal stem/stromal cells (MSC) are promising candidates for use in cell-based therapies. In most cases, therapeutic response appears to be cell-dose ...

Chondrogenic Differentiation Induction of Adipose-derived Stem Cells by Centrifugal Gravity

Santrifüj Yerçekimi ile adipoz kaynaklı Kök Hücrelerin kondrojenik Farklılaşma İndüksiyon

Impaired cartilage cannot heal naturally. Currently, the most advanced therapy for defects in cartilage is the transplantation of chondrocytes ...

Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization

Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization

Fare Endometrial stromal ve epitelyal hücrelerin izolasyonu için<em&gt; İn Vitro</em&gt; sidualizasyonu

Decidualization is a progesterone-dependent differentiation process of endometrial stromal cells and is a prerequisite for successful embryo implantation. ...

Isolation and Characterization of Mesenchymal Stromal Cells from Human Umbilical Cord and Fetal Placenta

İnsan göbek bağı ve fetal plasentadan Mezenkimal Stromal Hücre İzolasyonu ve Vasıflandırılması

The human umbilical cord (UC) and placenta are non-invasive, primitive and abundant sources of mesenchymal stromal cells (MSCs) that have increasingly ...

In Vivo Tracking of Human Adipose-derived Mesenchymal Stem Cells in a Rat Knee Osteoarthritis Model with Fluorescent Lipophilic Membrane Dye

In Vivo Tracking of Human Adipose-derived Mesenchymal Stem Cells in a Rat Knee Osteoarthritis Model with Fluorescent Lipophilic Membrane Dye

Vivo İnsan yağ elde edilen Mezenkimal kök hücrelerin sıçan diz osteoartrit modelinde floresan lipofilik membran boya ile izleme

In order to support the clinical application of human adipose-derived mesenchymal stem cell (haMSC) therapy for knee osteoarthritis (KOA), we examined the ...

Isolation of Human Endometrial Stromal Cells for In Vitro Decidualization

Isolation of Human Endometrial Stromal Cells for In Vitro Decidualization

Vitro Decidualization için insan endometrium Stromal hücre izolasyon

The differentiation of human endometrial stromal cells (HESC) from fibroblast-like appearance into secretory decidua is a transformation required for ...