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Isolation, Phenotyping, and Differentiation of Umbilical Cord Mesenchymal Stem Cells (UCMSCs)

عزل الخلايا الجذعية المتوسطة من نسيج الحبل السري و تنميطها و تمايزها

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 Publication date 2012
  fields Pharmacy
and research's language is العربية
 Created by Shamra Editor




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This research aims at establishing and testing protocols for isolation, in vitro proliferation, phenotyping, and differentiation of MSCs embedded in umbilical cord tissues of Syrian newborns. MSCs were isolated from two caesarian births depending on their adherence characteristic on plastic surfaces, and cells were cultured in MSC medium for their growth and proliferation. Cell phenotyping was performed by flow cytometry using fluorescent monoclonal antibodies specific to MSCs’ surface markers. Cultured cells were passed several times and a portion of these cells was cryopreserved in liquid nitrogen, and cell viability was assessed. Differentiation of these MSCs into adipocytes was conducted using culture medium Indomethacin and hydrocortisone.


Artificial intelligence review:
Research summary
تهدف هذه الدراسة إلى عزل الخلايا الجذعية المتوسطة (MSCs) من نسيج الحبل السري لمواليد سوريين، وتكثيرها في أوساط استنبات ملائمة، وتحريض تمايزها. تم عزل الخلايا من عينتين من الحبل السري باستخدام خاصية التصاق هذه الخلايا بالسطوح البلاستيكية. تم استنبات الخلايا في وسط مغذ خاص يسمح بنموها وتكاثرها، وتم تحديد خصائصها باستخدام تقنية الجريان الخلوي. أظهرت الخلايا المعزولة تعبيراً إيجابياً عن المستضدات السطحية المميزة للخلايا الجذعية المتوسطة مثل CD90 وCD105 وCD13، وسلبية أو ضعيفة التعبير عن المستضدات المميزة للخلايا الجذعية المولدة للدم مثل CD34. تم تحريض تمايز الخلايا إلى خلايا شحمية باستخدام الإندوميتاسين والهيدروكورتيزون، وأظهرت الخلايا المتمايزة تغييرات في الشكل وظهور قطيرات دهنية تلونت بأحمر السودان. تم حفظ الخلايا بالتجميد في الأزوت السائل مع الحفاظ على عيوشية عالية بلغت حوالي 90%. تعتبر هذه الدراسة الأولى من نوعها في سوريا لعزل الخلايا الجذعية المتوسطة من نسيج الحبل السري، وتؤسس لمزيد من البحوث المستقبلية في مجال الطب التجديدي وعلاج العديد من الأمراض.
Critical review
تعد هذه الدراسة خطوة مهمة في مجال البحث العلمي في سوريا، حيث تساهم في تأسيس بنوك لدماء وأنسجة الحبل السري. ومع ذلك، يمكن توجيه بعض الانتقادات البناءة لتحسين البحث في المستقبل. أولاً، كان من الممكن توسيع نطاق الدراسة لتشمل عدد أكبر من العينات لتأكيد النتائج بشكل أكبر. ثانياً، لم يتم التطرق بشكل كافٍ إلى التحديات التي قد تواجه عملية العزل والتكثير في بيئات مختلفة. أخيراً، كان من الممكن تقديم تفاصيل أكثر حول الإجراءات المتبعة لضمان عدم تلوث العينات، خاصة وأن العينات المأخوذة من الولادات الطبيعية أظهرت تلوثاً جرثومياً.
Questions related to the research
  1. ما هو الهدف الرئيسي من هذه الدراسة؟

    الهدف الرئيسي هو عزل وتكثير الخلايا الجذعية المتوسطة من نسيج الحبل السري لمواليد سوريين وتحريض تمايزها.

  2. ما هي المستضدات السطحية التي أظهرت الخلايا المعزولة تعبيراً إيجابياً عنها؟

    أظهرت الخلايا تعبيراً إيجابياً عن المستضدات السطحية CD90 وCD105 وCD13.

  3. ما هي المواد المستخدمة لتحريض تمايز الخلايا إلى خلايا شحمية؟

    تم استخدام الإندوميتاسين والهيدروكورتيزون لتحريض تمايز الخلايا إلى خلايا شحمية.

  4. ما هي نسبة عيوشية الخلايا بعد حفظها بالتجميد؟

    بلغت نسبة عيوشية الخلايا حوالي 90% بعد حفظها بالتجميد في الأزوت السائل.


References used
Cognet, P.A., and Minguell J. J., 1999. Phenotypical and functional properties of human bone marrow mesenchymal progenitor cells. J. Cell Physiol; V.181, pp. 67-73
Meirelles, L.D.S., Caplan, A.I., and Nardi, N., 2008. In search of the in vivo identity of mesenchymal stem cells. Stem Cells; V.26, pp. 2287–2299
Haniffa, M.A., Collin, M.P., Buckley, C.D., and Dazzi, F., 2009. Mesenchymal stem cells: the fibroblasts’ new clothes? Haematologica; V.94, pp. 258-263
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Stem cells have unique capability to differentiate into many cell types that can normally replace the loss in some cells of the body due to tissue injury. Umbilical cord blood (UCB) and umbilical cord (UC) are the two main sources for hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs), respectively, which constitutes the basis for stem cell banks that have been established worldwide and very recently in Syria. Research in our region has mainly focused on cell storage and freezing protocols, and only few studies were conducted to prove the ability of the stored cells to differentiate into their destined lineages. This study aimed to test the potential of cryopreserved MSCs isolated from an umbilical cord taken from new delivery at Maternity University Hospital in Damascus, to differentiate into various types of cells in response to growth and induction factors specific to cell lineages.
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