Sains Malaysiana 39(2)(2010): 305–313

 

Keupayaan Pembezaan Tiga Jenis Sel Primitif daripada Hasil Perbezaan Tempoh Proliferasi Darah Mencit

(Potential Differentiation of Three Types of Primitive Cells Originated from Different Proliferation Terms of Mouse Blood)

 

Intan Zarina Zainol Abidin & Shahrul Hisham Zainal Ariffin*

Pusat Pengajian Biosains dan Bioteknologi, Fakulti Sains dan Teknologi

Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, Malaysia

 

Zaidah Zainal Ariffin

Jabatan Mikrobiologi, Jakulti Sains Gunaan, Universiti Teknologi MARA

40450 Shah Alam, Selangor, Malaysia

 

Rohaya Megat Abdul Wahab

Jabatan Ortodontik, Fakulti Pergigian, Universiti Kebangsaan Malaysia

Jalan Raja Muda Aziz, 50300 Kuala Lumpur, Malaysia

 

Diserahkan: 24 Mac 2009 / Diterima: 13 Julai 2009

 

ABSTRAK

 

Kajian ini dilakukan bagi melihat keupayaan sel mononukleus sistem darah pusat membeza kepada sel osteoblas dan osteoklas secara in vitro bagi tiga tempoh proliferasi yang berbeza. Sel mononukleus sistem darah pusat dikulturkan di dalam medium pemilihan proliferasi bagi tiga tempoh proliferasi yang berbeza iaitu jangkamasa pendek (5 hari), sederhana (15 hari) dan panjang (30 hari). Keupayaan sel mononukleus untuk membeza kepada sel osteoblas dan osteoklas seterusnya diperhatikan pada setiap jenis sel ini. Medium proliferasi ditambah dengan faktor pembezaan asid askorbik dan β-gliserofosfat bagi membezakan sel mononukleus kepada sel osteoblas. Bagi pembezaan sel osteoklas pula, RANKL dan M-CSF ditambah ke dalam medium proliferasi. Bagi kawalan, sel yang sama digunakan tanpa penambahan faktor pembezaan. Viabiliti sel yang membeza daripada sel jangkamasa pendek, sederhana dan panjang menunjukkan sel-sel tersebut berupaya untuk bermandiri tanpa sebarang peningkatan yang signifikan sehingga 10 dan 14 hari dengan kehadiran faktor-faktor pembezaan tertentu di dalam medium pembezaan masing-masing. Analisis biokimia ke atas aktiviti alkali fosfatase (ALP) dan asid fosfatase rintang tartarat (TRAP) menunjukkan peningkatan yang signifikan (p<0.05) apabila dikulturkan di dalam medium pembezaan masing-masing. Kesimpulannya, keupayaan sel primitif untuk membeza kepada sel osteoblas dan osteoklas matang adalah hampir sama bagi ketiga-tiga jenis jangkamasa proliferasi tetapi mempunyai kadar proliferasi yang berlainan iaitu 0.37, 0.55 dan 0.72 pembahagian/hari masing-masing bagi sel jangkamasa pendek, sederhana dan panjang. Sel mononukleus yang diasingkan daripada darah periferi ini sangat primitif kerana berpotensi untuk membeza kepada dua jenis sel matang yang berasal daripada sel stem yang berbeza, justeru boleh dikategorikan sebagai sel stem multipoten.

 

Kata kunci: Darah periferi; mencit; osteoblas; osteoklas; sel stem

 

                                                              ABSTRACT           

 

The aim of this study was to differentiate central blood system mononucleated cells in vitro into osteoblast and osteoclast cells for three different proliferation terms of cells. The mononucleated cells were cultured in a selective proliferation medium for three different proliferation terms, i.e., short (5 days), medium (15 days) and long term (30 days) prior to analysis of osteoblast and osteoclast cells’ differentiation potentialities. The proliferation medium was then supplemented with differentiation factors, i.e., ascorbic acid and β-glycerophosphate to differentiate mononucleated cells into osteoblast cells. For osteoclast assay, RANKL and M-CSF were added into proliferation medium. For control, the same cells were used without supplementation of respective differentiation factors. The viability of differentiated cells from short, medium and long types of cells showed that they were able to survive until 10 to 14 days in the presence of respective differentiation factors without significant increased in the specific differentiation medium. Biochemical analyses on both alkaline phosphatase (ALP) and tartrate resistant acid phosphatase (TRAP) activities were significantly increased (p<0.05) once cultured in their respective differentiation medium. In conclusion, the three types of primitive cells have the same potentiality to differentiate into mature osteoblast and osteoclast cells even though the proliferation rates are different, i.e. 0.37, 0.55 and 0.72 division/day for short, medium and long term cells respectively. Mononucleated cells isolated from peripheral blood are primitive enough to differentiate into two distinct types of mature cells which originated from two different stem cells lineage hence can be categorized as multipotent stem cells.

 

Keywords: Mice; osteoblast; osteoclast; peripheral blood; stem cells

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*Pengarang untuk surat-menyurat; email: hisham@ukm.my