Sains Malaysiana 47(10)(2018): 2369–2379

http://dx.doi.org/10.17576/jsm-2018-4710-13

 

Chondrogenesis of Adipose-Derived Stem Cells with Chondrocytes in Low Serum towards Clinical Application

(Sel Asal Kondrogenesis Terbitan Adipos dengan Kondrosit dalam Serum Rendah untuk Aplikasi Klinikal)

 

ADILA A HAMID^1*, SATISH VAARMAN JEYABALAN¹, ALEZA OMAR1, NIK ZATTIL HANAN MOHD YASSIN¹, WONG TZENG LIN¹, LIAU LING LING¹, NUR AZURAH ABDUL GHANI¹, ANEEZA KHAIRIYAH WAN HAMIZAN¹ & CHUA KIEN HUI¹

 

¹Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latiff, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

 

²Department of Obstetrics and Gynaecology, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latiff, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

 

³Department of Otorhinolaryngology, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latiff, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

 

Diserahkan: 22 Mac 2018/Diterima: 5 Jun 2018

 

 

ABSTRACT

Currently, fetal bovine serum (FBS) have been widely use in culture media to promote human cell proliferation. However, the usage of FBS for cell therapy in clinical application was associated with the risk of viral and prion transmission as well as immune rejection. To provide an option for this risk, this study was conducted to determine the effect of adipose derived stem cells (ADSCs) co-culture with chondrocyte in promoting cell proliferation and chondrogenesis toward FBS free condition. ADSCs co-cultured with chondrocyte at the ratio of 1:1, 2:1 and 1:2 were tested. Cell morphology changes, cell proliferation and gene expression level of stemness (Oct4, FGF-4, Nanog) and chondrogenic (Collagen Type II, ACP) were assessed. The results showed ADSCs in all co-culture groups changed morphology from fibroblastic spindle to polygonal shape which resembled chondrocytes. The morphological changes were accompanied with increased expression of chondrogenic genes; denoted chondrogenesis process. While maintaining expression of stemness genes indicated continuation of cell proliferation. From the three co-culture groups tested; ADSCs and chondrocytes (1:1 ratio) have been shown to exert better effects in promoting cell proliferation and chondrogenesis. In conclusion, ADSCs could replace FBS to grow sufficient number of chondrogenic cells to repair cartilage injury in the near future. Further in vivo study should be performed to test the effectiveness of this co-culture technique in cartilage injury repair.

 

Keywords: Cartilage; chondrocyte; clinical application; serum; stem cell

 

ABSTRAK

Pada masa kini, serum fetus bovin (FBS) telah digunakan secara meluas di dalam media kultur sebagai faktor penggalak pertumbuhan sel. Namun begitu, penggunaan FBS untuk terapi sel secara klinikal dikaitkan dengan risiko transmisi virus dan prion serta tolakan imun. Bagi menangani risiko ini, kajian ini dijalankan bagi menentukan kesan ko-kultur sel stem adipos (ADSCs) dan kondrosit terhadap proliferasi sel dan kondrogenesis kearah kondisi pengkulturan bebas FBS. ADSCs dikultur bersama kondrosit dengan nisbah 1:1, 1:2 dan 2:1 telah diuji. Perubahan morfologi sel, proliferasi sel dan ekspresi gen kesteman (Oct4, FGF-4, Nanog) dan gen kondrogenik (kolagen jenis II dan ACP) telah dinilai. Keputusan kajian mendapati ADSCs dalam semua kumpulan ko-kultur berubah morfologi daripada bentuk fibroblas memanjang kepada bentuk poligon yang menyerupai kondrosit. Perubahan bentuk ini turut disertai dengan ekpresi gen kondrogenik yang meningkat, ini menandakan proses kondrogenesis telah berlaku. Sementara itu, ekspresi gen kesteman memberi penunjuk bahawa proliferasi sel dapat diteruskan. Daripada tiga kumpulan kajian nisbah ko-kultur yang dikaji; ADSCs bersama kondrosit (1:1 ratio) telah menunjukkan kesan yang lebih ketara dalam menggalakkan proliferasi sel dan kondrogenesis. Kesimpulannya, ADSCs berpotensi menggantikan FBS sebagai faktor pertumbuhan bagi menyediakan jumlah kondrosit yang mencukupi untuk merawat kecederaan rawan pada masa hadapan. Kajian seterusnya di dalam model in vivo perlu dijalankan bagi menguji keberkesanan teknik ko-kultur ini bagi merawat kecederaan rawan.

 

Kata kunci: Kondrosit; penggunaan klinikal; rawan; sel asal; serum

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