Name: preparation of pooled human platelet lysate (phpl) as an efficient supplement for animal serum-free human stem cell cultures
Uploaded: 2009-10-30T07:00:00
Description: Watch this Scientific Journal Video about Preparation of Pooled Human Platelet Lysate pHPL as an Efficient Supplement for Animal Serum-Free Human Stem Cell Cultures at JoVE.com

وقد تم دعم هذا العمل من قبل مؤسسة الأبحاث النمساوية (FWF ، منح N211 - NAN إلى DS) وتعزيز البحوث النمساوية وكالة (FFG ، ومنحة لN200 DS). الكتاب نشكر كلوديا رابط للحصول على مساعدة فنية ممتازة وفاريل مونيكا لتحرير اللغوي.

1. بدءا المواد تبدأ الصفيحات الغنية وحدات (PRP) البلازما التي أعدتها فصادة الكريات أو مشتقة من المعاطف الشهباء. 2. عقم الاختيار للعقم الاختيار أخذ عينة من 20 مل ...

<ol>
	<li>Nurden, A. T., Nurden, P., Sanchez, M., Andia, I., Anitua, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Platelets+and+wound+healing.">Platelets and wound healing.</a> Front Biosci. 13, 3532-3548 (2008).</li><li>Barrientos, S., Stojadinovic, O., Golinko, M. S., Brem, H., Tomic-Canic, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growth+factors+and+cytokines+in+wound+healing.">Growth factors and cytokines in wound healing.</a> Wound Repair Regen. 16, 585-601 (2008).</li><li>Giacco, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thrombin-activated+platelets+induce+proliferation+of+human+skin+fibroblasts+by+stimulating+autocrine+production+of+insulin-like+growth+factor-1.">Thrombin-activated platelets induce proliferation of human skin fibroblasts by stimulating autocrine production of insulin-like growth factor-1.</a> FASEB J. 20, 2402-2404 (2006).</li><li>Kocaoemer, A., Kern, S., Kluter, H., Bieback, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+AB+serum+and+thrombin-activated+platelet-rich+plasma+are+suitable+alternatives+to+fetal+calf+serum+for+the+expansion+of+mesenchymal+stem+cells+from+adipose+tissue.">Human AB serum and thrombin-activated platelet-rich plasma are suitable alternatives to fetal calf serum for the expansion of mesenchymal stem cells from adipose tissue.</a> Stem Cells. 25, 1270-1278 (2007).</li><li>Kakudo, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proliferation-promoting+effect+of+platelet-rich+plasma+on+human+adipose-derived+stem+cells+and+human+dermal+fibroblasts.">Proliferation-promoting effect of platelet-rich plasma on human adipose-derived stem cells and human dermal fibroblasts.</a> Plast Reconstr Surg. 122, 1352-1360 (2008).</li><li>Doucet, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Platelet+lysates+promote+mesenchymal+stem+cell+expansion:+A+safety+substitute+for+animal+serum+in+cell-based+therapy+applications.">Platelet lysates promote mesenchymal stem cell expansion: A safety substitute for animal serum in cell-based therapy applications.</a> J Cell Physiol. 205, 228-236 (2005).</li><li>Schallmoser, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+platelet+lysate+can+replace+fetal+bovine+serum+for+clinical-scale+expansion+of+functional+mesenchymal+stromal+cells.">Human platelet lysate can replace fetal bovine serum for clinical-scale expansion of functional mesenchymal stromal cells.</a> Transfusion. 47, 1436-1446 (2007).</li><li>Reinisch, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Humanized+system+to+propagate+cord+blood-derived+multipotent+mesenchymal+stromal+cells+for+clinical+application.">Humanized system to propagate cord blood-derived multipotent mesenchymal stromal cells for clinical application.</a> Regen Med. 2, 371-382 (2007).</li><li>Schallmoser, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+large-scale+expansion+of+functional+mesenchymal+stem+cells+from+unmanipulated+bone+marrow+without+animal+serum+Tissue+Eng+Part.">Rapid large-scale expansion of functional mesenchymal stem cells from unmanipulated bone marrow without animal serum Tissue Eng Part.</a> C Methods. 14, 185-196 (2008).</li><li>Lange, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accelerated+and+safe+expansion+of+human+mesenchymal+stromal+cells+in+animal+serum-free+medium+for+transplantation+and+regenerative+medicine.">Accelerated and safe expansion of human mesenchymal stromal cells in animal serum-free medium for transplantation and regenerative medicine.</a> J Cell Physiol. 213, 18-26 (2007).</li><li>Reinisch, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Humanized+large-scale+expanded+endothelial+colony-forming+cells+function+in+vitro+and+in+vivo.">Humanized large-scale expanded endothelial colony-forming cells function in vitro and in vivo.</a> Blood. 113, 6716-6725 (2009).</li><li>Bernardo, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimization+of+in+vitro+expansion+of+human+multipotent+mesenchymal+stromal+cells+for+cell-therapy+approaches:+further+insights+in+the+search+for+a+fetal+calf+serum+substitute.">Optimization of in vitro expansion of human multipotent mesenchymal stromal cells for cell-therapy approaches: further insights in the search for a fetal calf serum substitute.</a> J Cell Physiol. 211, 121-130 (2007).</li><li>Carrancio, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimization+of+mesenchymal+stem+cell+expansion+procedures+by+cell+separation+and+culture+conditions+modification.">Optimization of mesenchymal stem cell expansion procedures by cell separation and culture conditions modification.</a> Exp Hematol. 36, 1014-1021 (2008).</li><li>Muller, 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=Platelet+lysate+as+a+serum+substitute+for+2D+static+and+3D+perfusion+culture+of+stromal+vascular+fraction+cells+from+human+adipose+tissue.">Platelet lysate as a serum substitute for 2D static and 3D perfusion culture of stromal vascular fraction cells from human adipose tissue.</a> Tissue Eng Part A. 15, 869-875 (2009).</li><li>Anitua, E., Sanchez, M., Orive, G., Andia, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+potential+impact+of+the+preparation+rich+in+growth+factors+(PRGF)+in+different+medical+fields.">The potential impact of the preparation rich in growth factors (PRGF) in different medical fields.</a> Biomaterials. 28, 4551-4560 (2007).</li><li>Anitua, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+insights+into+and+novel+applications+for+platelet-rich+fibrin+therapies.">New insights into and novel applications for platelet-rich fibrin therapies.</a> Trends Biotechnol. 24, 227-234 (2006).</li><li>Martinez-Zapata, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+and+safety+of+the+use+of+autologous+plasma+rich+in+platelets+for+tissue+regeneration:+a+systematic+review.">Efficacy and safety of the use of autologous plasma rich in platelets for tissue regeneration: a systematic review.</a> Transfusion. 49, 44-56 (2009).</li><li>Weibrich, G., Kleis, W. K., Hafner, G., Hitzler, W. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growth+factor+levels+in+platelet-rich+plasma+and+correlations+with+donor+age,+sex,+and+platelet+count.">Growth factor levels in platelet-rich plasma and correlations with donor age, sex, and platelet count.</a> J Craniomaxillofac Surg. 30, 97-102 (2002).</li><li>Frechette, J. P., Martineau, I. &. a. m. p. ;. a. m. p., Gagnon, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Platelet-rich+plasmas:+growth+factor+content+and+roles+in+wound+healing.">Platelet-rich plasmas: growth factor content and roles in wound healing.</a> J Dent Res. 84, 434-439 (2005).</li><li>Seghatchian, J., Krailadsiri, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+platelet+storage+lesion.">The platelet storage lesion.</a> Transfus Med Rev. 11, 130-144 (1997).</li><li>Selvaggi, T. A., Walker, R. E., Fleisher, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+antibodies+to+fetal+calf+serum+with+arthus-like+reactions+in+human+immunodeficiency+virus-infected+patients+given+syngeneic+lymphocyte+infusions.">Development of antibodies to fetal calf serum with arthus-like reactions in human immunodeficiency virus-infected patients given syngeneic lymphocyte infusions.</a> Blood. 89, 776-779 (1997).</li><li>Tonti, G. A., Mannello, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+bone+marrow+to+therapeutic+applications:+different+behaviour+and+genetic/epigenetic+stability+during+mesenchymal+stem+cell+expansion+in+autologous+and+foetal+bovine+sera.">From bone marrow to therapeutic applications: different behaviour and genetic/epigenetic stability during mesenchymal stem cell expansion in autologous and foetal bovine sera.</a> Int J Dev Biol. 52, 1023-1032 (2008).</li><li>Bieback, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+Alternatives+to+Fetal+Bovine+Serum+for+the+Expansion+of+Mesenchymal+Stromal+Cells+from+Bone+Marrow.">Human Alternatives to Fetal Bovine Serum for the Expansion of Mesenchymal Stromal Cells from Bone Marrow.</a> Stem Cells. , (2009).</li><li>Rohde, E., Schallmoser, K., Bartmann, C., Reinisch, A., Gad, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=GMP-Compliant+Propagation+of+Human+Multipotent+Mesenchymal+Stromal+Cells.">GMP-Compliant Propagation of Human Multipotent Mesenchymal Stromal Cells.</a> Pharmaceutical Manufacturing Handbook: Regulations and Quality. , 97-115 (2008).</li></ol>

ربما في بعض المناطق يمكن الحصول على الصفائح الدموية البلازما الغنية (PRP) من الشهباء ، المعاطف يجري خلاف ذلك مضيعة للمنتج معبأة انتاج خلايا الدم الحمراء من تبرعات الدم اختبار (الشكل 1). على النحو الأمثل ، يتم استخدام PRP على الفور لإعداد المزيد من pHPL ، كما في الصفائح الدمو...

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Double bag (2 x 3.5L)</td>
<td>&nbsp;</td>
<td>MacoPharma</td>
<td>VDL 8000XQ</td>
<td>originally for ascites puncture</td>
</tr>
<tr>
<td>50 mL Falcon tube</td>
<td>&nbsp;</td>
<td>Falcon BD</td>
<td>2098</td>
<td>&nbsp;</td>
<tr>
<td>50 mL Stripettes</td>
<td>&nbsp;</td>
<td>Costar</td>
<td>4501</td>
<td>&nbsp;</td>
<tr>
<td>Plasma bags (600mL)</td>
<td>&nbsp;</td>
<td>Baxter</td>
<td>R4R2021</td>
<td>&nbsp;</td>
<tr>
<td>Sterile tubing welder</td>
<td>&nbsp;</td>
<td>Terumo</td>
<td>TSCD-II</td>
<td>&nbsp;</td>
<tr>
<td>Welding equipment </td>
<td>&nbsp;</td>
<td>Fresenius Kabi</td>
<td>Compo Seal</td>
<td>&nbsp;</td>
<tr>
<td>Water bath</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Sterile scissors</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Clamps</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Centrifuge</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

preparation of pooled human platelet lysate (phpl) as an efficient supplement for animal serum-free human stem cell cultures

وقد تبين أن الصفيحات عوامل النمو المشتقة لتحفيز تكاثر الخلايا بكفاءة<em&gt; في الجسم الحي</em<sup&gt; 1،2</sup&gt; و<em&gt; في المختبر</em&gt;. وقد أبلغ هذا التأثير لخلايا انسجة الوسيطة (MSCS) ، الليفية والخلايا البطانية التي تشكل مع مستعمرة الصفائح تفعيلها من خلال ثرومبين<sup&gt; 3-5</sup&gt; أو بواسطة lysed تجميد / الذوبان دورات<sup&gt; 14/06</sup&gt; قبل إضافة releasate الصفيحات إلى الخلية متوسطة الثقافة. وقد تم بالفعل تأثير التغذية من عوامل النمو المشتقة الصفيحات اختبارها في عدة محاكمات لهندسة الأنسجة والعلاج التجديدي.<sup&gt; 1،15-17</supيعتبر&gt; كفاءة متفاوتة على الأقل في جزء منه نتيجة لتركيزات مختلفة من عوامل نمو فردي<sup&gt; 18،19</sup&gt; والنقص الحالي في بروتوكولات موحدة لإعداد الصفائح الدموية.<sup&gt; 15،16</sup&gt; يعرض هذا البروتوكول إجراء العملية لتوليد مجموعة من الصفائح الدموية lysate الإنسان (pHPL) المستمدة من البلازما التي تنتج عادة غنية الصفيحات (PRP) من 40-50 التبرع بالدم واحد. بواسطة دورات تجميد / الذوبان عدة معطوبة الأغشية والصفائح الدموية يتم الافراج بكفاءة عوامل النمو في البلازما. أخيرا ، تتم إزالة أجزاء الصفائح الدموية عن طريق الطرد المركزي لتجنب تشكيل التجميعية واسعة وتستنزف مستضدات محتملة. تنفيذ pHPL في البروتوكولات القياسية الثقافة تمثل أداة واعدة لمزيد من تطوير علاجات الخلية نشر في البروتين الحيواني خالية من النظام.

Watch this Scientific Journal Video about Preparation of Pooled Human Platelet Lysate pHPL as an Efficient Supplement for Animal Serum-Free Human Stem Cell Cultures at JoVE.com

Preparation of Pooled Human Platelet Lysate pHPL as an Efficient Supplement for Animal Serum-Free Human Stem Cell Cultures

lysate الصفيحات الإنسان هو مصدر غني من عوامل النمو وقوية في ملحق ثقافة الخلية. يعرض هذا البروتوكول عملية إعداد مجموعة كبيرة من الصفائح الدموية lysate الإنسان بدءا من البلازما الغنية الصفائح الدموية ، وأداء العديد من تجميد أذاب الدورات واستنزاف شظايا الصفائح الدموية.

إعداد Lysate مجمع صفائح الدم البشرية (pHPL) باعتبارها الملحق لمصل فعال خالية من الإنسان الحيوان الثقافات الخلايا الجذعية

Platelet derived growth factors have been shown to stimulate cell proliferation efficiently in vivo1,2 and in vitro. This effect has been reported for ...

Research

JoVE Journal

Biology

Preparation of Pooled Human Platelet Lysate (pHPL) as an Efficient Supplement for Animal Serum-Free Human Stem Cell Cultures

Mouse Epidermal Neural Crest Stem Cell (EPI-NCSC) Cultures

Mouse Epidermal Neural Crest Stem Cell EPI-NCSC Cultures

الماوس البشرة الجذعية العصبية خلية كريست (EPI - NCSC) الثقافات

EPI-NCSC are remnants of the embryonic neural crest in an adult location, the bulge of hair follicles. They are multipotent stem cells that have the ...

Preparation of Aplysia Sensory-motor Neuronal Cell Cultures

Preparation of Aplysia Sensory-motor Neuronal Cell Cultures

إعداد Aplysia الحسية والحركية الثقافات الخلية متعلق بالخلايا العصبية

The nervous system of the marine mollusk Aplysia californica is relatively simple, consisting of approximately 20,000 neurons.  The neurons are large (up ...

Isolation and Animal Serum Free Expansion of Human Umbilical Cord Derived Mesenchymal Stromal Cells (MSCs) and Endothelial Colony Forming Progenitor Cells (ECFCs)

Isolation and Animal Serum Free Expansion of Human Umbilical Cord Derived Mesenchymal Stromal Cells MSCs and Endothelial Colony Forming Progenitor Cells ECFCs

العزلة والحيوان التوسع الحرة المصل الحبل السري الإنسان المستمدة الخلايا اللحمية الوسيطة (MSCS) وغشائي خلايا مستعمرة السلف التشكيل (ECFCs)

The umbilical cord is a rich source for progenitor cells with high proliferative potential including mesenchymal stromal cells (also termed mesenchymal ...

Differentiation of Embryonic Stem Cells into Oligodendrocyte Precursors

تمايز الخلايا الجذعية الجنينية في السلائف Oligodendrocyte

Oligodendrocytes are the myelinating cells of the central nervous system. For regenerative cell therapy in demyelinating diseases, there is significant ...

Xenotransplantation of Human Stem Cells into the Chicken Embryo

زرع الأعضاء من الخلايا الجذعية البشرية إلى الأجنة الدجاج

The chicken embryo is a classical animal model for studying normal embryonic and fetal development and for xenotransplantation experiments to study the ...

Efficient Derivation of Human Cardiac Precursors and Cardiomyocytes from Pluripotent Human Embryonic Stem Cells with Small Molecule Induction

كفاءة اشتقاق السلائف القلب البشري وافرة القوة العضلية من خلايا المنشأ الجنينية البشرية التعريفي مع جزيء صغير

To date, the lack of a suitable human cardiac cell source has been the major setback in regenerating the human myocardium, either by cell-based ...

Rapid Fibroblast Removal from High Density Human Embryonic Stem Cell Cultures

إزالة الخلايا الليفية السريع من الكثافة البشرية العالية الجنينية الخلايا الجذعية الثقافات

Mouse embryonic fibroblasts (MEFs) were used to establish human embryonic stem cells (hESCs) cultures after blastocyst isolation1. This feeder system ...

Na&iuml;ve Adult Stem Cells Isolation from Primary Human Fibroblast Cultures

Na&#239;ve Adult Stem Cells Isolation from Primary Human Fibroblast Cultures

السذاجة الكبار الخلايا الجذعية عزل من المرحلة الابتدائية الثقافات الخلايا الليفية الإنسان

Over the last decade, several adult stem cell  populations have been identified in human skin 1-4.  The isolation of multipotent adult dermal precursors ...

Efficient Generation Human Induced Pluripotent Stem Cells from Human Somatic Cells with Sendai-virus

كفاءة الجيل المستحثة الإنسان افرة القوة الخلايا الجذعية من الخلايا الجسدية البشرية مع سينداي الفيروسات

A few years ago, the establishment of human induced pluripotent stem cells (iPSCs) ushered in a new era in biomedicine. Potential uses of human iPSCs ...

Alternative Cultures for Human Pluripotent Stem Cell Production, Maintenance, and Genetic Analysis

الثقافات البديلة لافرة القوة البشرية الجذعية إنتاج الخلية، الصيانة، والتحليل الوراثي

Human pluripotent stem cells (hPSCs) hold great promise for regenerative medicine and biopharmaceutical applications. Currently, optimal culture and ...