Tehran University Medical Journal
مجله دانشکده پزشکی، دانشگاه علوم پزشکی تهران
Tehran Univ Med J
Medical Sciences
http://tumj.tums.ac.ir
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admin
1683-1764
1735-7322
10.18869/acadpub.tumj
000
000
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fa
jalali
1391
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gregorian
2012
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online
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fulltext
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قابلیت تمایز سلولهای بنیادی جنین انسان (Royan H5) به سلولهای همانژیوبلاست در شرایط آزمایشگاهی
In-vitro differentiation of human embryonic stem cells into hemangioblasts
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<strong><span style="font-size: 10pt">زمینه و هدف: </span></strong><span style="font-size: 10pt">قابلیت تمایز سلولهای بنیادی جنین انسان بهعنوان سلولهای پرتوان، به انواع سلولهای مختلف از جمله سلولهای عصبی، قلبی و کبدی بررسی شده است. در این مطالعه با توجه به توانمندی سلولهای بنیادی جنین انسان </span><span style="font-size: 8pt">(</span><span style="font-size: 7pt">R</span><span style="font-size: 8pt">oyan </span><span style="font-size: 7pt">H5</span><span style="font-size: 8pt">)</span><span style="font-size: 10pt"> <span lang="FA">که در پژوهشگاه رویان بهصورت رده سلولی تهیه شده، بررسی قابلیت تمایز این سلولها به سلولهای خونی مورد توجه قرار گرفت. هدف مطالعه بررسی توانمندی سلولهای بنیادی جنین انسان<span> </span><span> </span></span></span><span style="font-size: 8pt">(</span><span style="font-size: 7pt">R</span><span style="font-size: 8pt">oyan </span><span style="font-size: 7pt">H5</span><span style="font-size: 8pt">)</span><span style="font-size: 10pt"> در تمایز به سلولهای همانژیوبلاست بود.</span><span style="font-size: 9pt"> </span></w:donotpromoteqf></w:validateagainstschemas></w:punctuationkerning></w:trackformatting></w:trackmoves></p>
<p dir="rtl" style="text-align: justify"><strong><span style="font-size: 10pt">روش بررسی:</span><em><span style="font-size: 10pt"> </span></em></strong><a name="OLE_LINK1"><span style="font-size: 10pt">سلولهای بنیادی جنین انسان بهصورت سوسپانسیون در محیط کامل </span><span style="font-size: 7pt">DMEM/F12</span><span style="font-size: 10pt"> و در حضور (</span><span style="font-size: 8pt">ngm</span><span style="font-size: 7pt">L</span><span style="font-size: 10pt">100) </span><span style="font-size: 8pt">b</span><span style="font-size: 7pt">FGF</span><span style="font-size: 10pt"> کشت و تکثیر شدند </span></a><span style="font-size: 10pt">و در روز هشت در حضور ترکیبات القاکننده به سلولهای بلاست تمایز داده شدند. شاخصهای سلولهای بلاست </span><span style="font-size: 7pt">KDR</span><span style="font-size: 10pt">، </span><span style="font-size: 7pt">CD31</span><span style="font-size: 10pt"> و </span><span style="font-size: 7pt">CD34</span><span style="font-size: 10pt"> توسط فلوسیتومتری و بیان ژنهای </span><span style="font-size: 7pt">TAL-1</span><span style="font-size: 10pt">، </span><span style="font-size: 8pt">Runx-</span><span style="font-size: 7pt">1</span><span style="font-size: 10pt"> و </span><span style="font-size: 7pt">CD34</span><span style="font-size: 10pt"> توسط </span><span style="font-size: 7pt">Q</span><span style="font-size: 8pt">uantitative </span><span style="font-size: 7pt">R</span><span style="font-size: 8pt">eal </span><span style="font-size: 7pt">T</span><span style="font-size: 8pt">ime-</span><span style="font-size: 7pt">PCR</span><span style="font-size: 10pt"> نشان داده شد و توان کلونیزایی بلاستها </span><span style="font-size: 8pt">(Colony forming unit-assay)</span><span style="font-size: 10pt"> <span lang="AR-SA">در محیط حاوی متیل سلولز ارزیابی شد.</span></span><strong><span style="font-size: 10pt"></span></strong></p>
<p dir="rtl" style="text-align: justify"><strong><span style="font-size: 10pt"></span></strong><strong><span style="font-size: 10pt">یافتهها:</span></strong><span style="font-size: 10pt"> </span><span style="font-size: 10pt">سلولهای بنیادی جنین انسان در طی تمایز به پیشسازهای خونی، جمعیتی از سلولهای (5/12%</span><span style="font-size: 8pt font-family: Symbol"><span>±</span></span><span style="font-size: 10pt">79) </span><span style="font-size: 7pt">KDR</span><sup><span style="font-size: 8pt">+</span></sup><span style="font-size: 10pt">، (8/2%</span><span style="font-size: 8pt">±</span><span style="font-size: 10pt">6/5) </span><span style="font-size: 7pt">CD31</span><sup><span style="font-size: 8pt">+</span></sup><span style="font-size: 8pt">-</span><span style="font-size: 7pt">CD34</span><sup><span style="font-size: 8pt">+</span></sup><span style="font-size: 10pt"> و (12/2%</span><span style="font-size: 8pt font-family: Symbol"><span>±</span></span><span style="font-size: 10pt">6) </span><span style="font-size: 7pt">KDR</span><sup><span style="font-size: 8pt">+</span></sup><span style="font-size: 8pt">-</span><span style="font-size: 7pt">CD31</span><sup><span style="font-size: 8pt">+</span></sup><span style="font-size: 10pt"> <span lang="FA">را بهوجود میآورند. افزایش معنیدار بیان ژنهای (01/0</span></span><span style="font-size: 7pt">P</span><span style="font-size: 8pt">≤</span><span style="font-size: 10pt">، 05/0</span><span style="font-size: 7pt">P</span><span style="font-size: 8pt">≤</span><span style="font-size: 10pt">) </span><span style="font-size: 7pt">TAL-1</span><span style="font-size: 8pt">, </span><span style="font-size: 7pt">RUNX-1</span><span style="font-size: 8pt">, </span><span style="font-size: 7pt">CD34</span><span style="font-size: 5pt"> </span><span style="font-size: 10pt"><span> </span><span lang="AR-SA">در کلونیهای </span></span><span style="font-size: 10pt">چهارده</span><span style="font-size: 10pt"> روزه شاهدی بر تشکیل سلولهای بلاست بود. همچنین کلونیهای شش روزه تولیدشده بر سطح ماتریژل و </span><span style="font-size: 10pt">با ویژگیهای شبه همانژیوبلاست در محیط حاوی متیل سلولز شبه کلونیهای مختلط خونی و سلولهای شبه اندوتلیال را ایجاد کردند.</span><strong><em><span style="font-size: 10pt"> </span></em></strong></p>
<p dir="rtl" style="text-align: justify"><strong><span style="font-size: 10pt">نتیجهگیری:</span><em><span style="font-size: 10pt"> </span></em></strong><span style="font-size: 10pt">سلولهای بنیادی جنینی </span><span style="font-size: 8pt">Royan </span><span style="font-size: 7pt">H5</span><span style="font-size: 10pt"> قادر به تولید سلولهای پیشساز خونی با توانمندی دوگانه در شرایط آزمایشگاهی میباشند ک</span><span style="font-size: 10pt">ه میتوانند کلونیهای شبیه به کلونیهای مختلط خونی و سلولهای شبه اندوتلیال را تولید کنند.</span></p>
<p style="text-align: justify"><strong>Background</strong>: Human embryonic stem cells (hESCs) are capable of self-renewal and large-scale expansion. They also have the capacity to differentiate into a variety of cell types including liver, cardiac and neuron cells. However, it is not yet clear whether hESCs can differentiate to hemangioblasts under in-vitro conditions. Hemangioblasts are bipotential progenitors that can generate hematopoietic lineages and endothelial cells. The aim of this study was to identify the potential of human Royan H5 embryonic stem cells in differentiating into hemangioblast cells.</p>
<p style="text-align: justify"><strong>Methods</strong>: HESCs were cultured at suspension system in DMEM/F12 supplemented with bFGF. 7-day old cells differentiated into blast cells under defined condition consisting of hematopoietic cytokines including BMP4, VEGF, etc. Blast cell markers kinase insert domain receptor (KDR), CD31, and CD34 were evaluated by flow cytometry and blast gene expressions (TAL-1, Runx-1 and CD34) were detected by qRT-PCR. Clonogenic assays were performed in semisolid medium by colony forming unit-assays.</p>
<p style="text-align: justify"><strong>Results</strong>: The hESCs (Royan H5) had the capacity of differentiating into hemangioblast cells. We could detect colonies that expressed 79%±12.5 KDR+, 5.6%±2.8 CD31+-CD34+ and 6%±2.12 KDR+-CD31+ on day 8 in the hESCs. Up-regulation of TAL-1, Runx-1 and CD34 occurred during hemangioblast commitment (P≤0.05 and P≤0.01, respectively). Moreover, hemangioblast cells generated mixed-type and endothelial-like colonies in semi-solid media.</p>
<p style="text-align: justify"><strong>Conclusion</strong>: Our results showed that hESCs (Royan H5) were able to differentiate into hemangioblasts under in-vitro conditions. The hemangioblasts had the potential to generate two non-adherent (Mixed-type) and adherent (endothelial-like) cell populations.</p>
سلولهای بنیادی جنینی انسانی,تمایز,کلونی همانژیوبلاست
Colony,differentiation,hemangioblast,human embryonic stem cells
141
149
http://tumj.tums.ac.ir/browse.php?a_code=A-10-4111-1&slc_lang=fa&sid=1
Ganji
Fatemeh
فاطمه
گنجی