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İnsan kemik iliği kaynaklı mezenkimal stromal hücrelerin kıkırdak ve kemik hücrelerine farklılaşma potansiyelinin değerlendirilmesi

Year 2007, Volume: 41 Issue: 4, 295 - 301, 05.12.2007

Abstract

Amaç: Bu çalışmada, sağlıklı insan kemik iliğinden ayrılarak çoğaltılan mezenkimal destekdoku (stromal) hücrelerinin (MSH) fiziksel özelliklerine ve yüzey antijenlerine göre tanımlanması, osteoblast ve kondroblast yönünde farklılaşma potansiyelinin incelenmesi amaçlandı.
Çalışma planı: Yaşları dört ay ile 18 yıl arasında değişen 10 kemik iliği transplantasyon vericisinden toplanan kemik iliğinin 1-3 ml’sinden gradiyent yöntemiyle mononükleer hücreler ayrıldı ve içinde %10 fetal dana serumu bulunan tüplerde kültüre edildi. Tabana yapışma gösteren fibroblastik görüntüde olan hücreler çoğaltıldı. Akım sitometri aygıtı ile hücrelerin yüzey antijenlerine göre immünfenotiplendirmesi yapıldı. Tanımlanan mezenkimal stromal hücrelerin osteoblast ve kondroblast uyarıcı medyumları kullanılarak kondroblast ve osteoblasta farklılaşması incelendi. Osteoblastlar Alizarin kırmızısı, kondroblastlar ise Alsiyan mavisi ile boyanarak mikrofotoğrafları çekildi.
Sonuçlar: Canlı-dışı (in vitro) çoğaltılan MSH’lerin biçim ve yapışma özellikleri yaşa bağlı farklılık göstermedi. Destekdoku kaynaklı olabilecek hücrelerde karakteristik CD105, CD44, CD166, CD29, CD90 ve CD73 antikorlarına karşı %90-99 arası pozitif sonuç, hematopoietik hücrelere özgü CD45, CD34, CD14 ve HLA-DR antikorlarına karşı negatif (%0-5) sonuç elde edildi. Hücre tipine özel besiyerlerinde çoğaltılan MSH’lerin, ilerleyen altkültürler (pasajlar) de dahil olmak üzere, tüm altkültürlerde (p1-15) osteoblast ve kondroblast hücrelerine farklılaştığı gözlendi.
Çıkarımlar: Mezenkimal destekdoku hücrelerin, kemik iliği aktarımı, kalıtsal hastalıklar ve organ hasarında; ortopedide kullanılması düşünülen biyomalzemelerin biyolojik etkilerinin canlı-dışı incelemesinde; kemik ve kıkırdak dokusundaki hasarlı bölgelerin tamirinde kullanılması mümkün görünmektedir.

Evaluation of differentiation potential of human bone marrow-derived mesenchymal stromal cells to cartilage and bone cells

Year 2007, Volume: 41 Issue: 4, 295 - 301, 05.12.2007

Abstract

Objectives: This study was designed to identify the characteristics and surface antigen properties of mesenchymal stromal cells (MSC) isolated and cultured from bone marrow of healthy subjects and to assess their differentiation potential to osteoblast and chondroblast lineage cells.
Methods: Mononuclear cells were isolated by density-gradient separation from 1-3 ml of bone marrow collected from 10 donors of bone transplantation aged between 4 months and 18 years. These mononuclear cells were cultured in flasks containing %10 fetal calf serum, which resulted in growth of fibroblast-like cells showing adhesion onto the culture flask. Physical properties of the cells were identified and flow cytometric immunophenotyping was performed to specify surface antigen properties. Differentiation of mesenchymal stromal cells in specific media to chondroblasts and osteoblasts was evaluated. Osteoblasts and chondroblasts were stained with Alizarin red and Alcian blue, respectively, and microphotographed.
Results: In vitro yield of MSCs showed no age-related differences in terms of morphological and adhesive properties. While cells of stromal origin showed strong positivity (90% to 99%) to characteristic CD105, CD44, CD166, CD29, CD90, and CD73 antibodies, hematopoietic cells remained negative (0% to 5%) to CD45, CD34, CD14, and HLA-DR antibodies. It was observed that MSCs produced in cell-specific media differentiated to osteoblasts and chondroblasts in all passages (p1-15) tested, including late passages.
Conclusion: It seems that the use of MSCs would provide promising treatment strategies in bone marrow transplantation, inherited diseases, and organ repair; in in vitro assessment of biological effects of biomaterials in orthopedics; and in repair of bone and cartilage injuries.

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Details

Primary Language English
Subjects Health Care Administration
Journal Section Original Article
Authors

Emine Kilic This is me

Taskin Ceyhan This is me

Duygu Cetinkaya This is me

Publication Date December 5, 2007
Published in Issue Year 2007 Volume: 41 Issue: 4

Cite

APA Kilic, E., Ceyhan, T., & Cetinkaya, D. (2007). Evaluation of differentiation potential of human bone marrow-derived mesenchymal stromal cells to cartilage and bone cells. Acta Orthopaedica Et Traumatologica Turcica, 41(4), 295-301.
AMA Kilic E, Ceyhan T, Cetinkaya D. Evaluation of differentiation potential of human bone marrow-derived mesenchymal stromal cells to cartilage and bone cells. Acta Orthopaedica et Traumatologica Turcica. December 2007;41(4):295-301.
Chicago Kilic, Emine, Taskin Ceyhan, and Duygu Cetinkaya. “Evaluation of Differentiation Potential of Human Bone Marrow-Derived Mesenchymal Stromal Cells to Cartilage and Bone Cells”. Acta Orthopaedica Et Traumatologica Turcica 41, no. 4 (December 2007): 295-301.
EndNote Kilic E, Ceyhan T, Cetinkaya D (December 1, 2007) Evaluation of differentiation potential of human bone marrow-derived mesenchymal stromal cells to cartilage and bone cells. Acta Orthopaedica et Traumatologica Turcica 41 4 295–301.
IEEE E. Kilic, T. Ceyhan, and D. Cetinkaya, “Evaluation of differentiation potential of human bone marrow-derived mesenchymal stromal cells to cartilage and bone cells”, Acta Orthopaedica et Traumatologica Turcica, vol. 41, no. 4, pp. 295–301, 2007.
ISNAD Kilic, Emine et al. “Evaluation of Differentiation Potential of Human Bone Marrow-Derived Mesenchymal Stromal Cells to Cartilage and Bone Cells”. Acta Orthopaedica et Traumatologica Turcica 41/4 (December 2007), 295-301.
JAMA Kilic E, Ceyhan T, Cetinkaya D. Evaluation of differentiation potential of human bone marrow-derived mesenchymal stromal cells to cartilage and bone cells. Acta Orthopaedica et Traumatologica Turcica. 2007;41:295–301.
MLA Kilic, Emine et al. “Evaluation of Differentiation Potential of Human Bone Marrow-Derived Mesenchymal Stromal Cells to Cartilage and Bone Cells”. Acta Orthopaedica Et Traumatologica Turcica, vol. 41, no. 4, 2007, pp. 295-01.
Vancouver Kilic E, Ceyhan T, Cetinkaya D. Evaluation of differentiation potential of human bone marrow-derived mesenchymal stromal cells to cartilage and bone cells. Acta Orthopaedica et Traumatologica Turcica. 2007;41(4):295-301.