Sains Malaysiana 50(7)(2021): 1987-1996

http://doi.org/10.17576/jsm-2021-5007-13

The Combination of bFGF and Hydrocortisone is a Better Alternative Compared to 5-Azacytidine for Cardiomyogenic Differentiation of Bone Marrow and Adipose Stem Cells

(Gabungan bFGF dan Hidrokortison adalah Alternatif yang Lebih Baik Berbanding dengan 5-Azasitidin bagi Perbezaan Kardomiogen Sumsum Tulang dan Sel Stem Adipos)

NADIAH SULAIMAN¹*, NUR QISYA AFIFAH VERONICA SAINIK^1,2, SHAMSUL BIN SULAIMAN¹, PEZHMAN HAFEZ¹, NG MIN HWEI¹ & RUSZYMAH BT HJ IDRUS^1,2

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

²Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latiff, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

Received: 27 April 2020/Accepted: 13 November 2020

ABSTRACT

Stem cells can be differentiated into cardiomyocytes by induction with 5-azacytidine (5-aza) but its carcinogenicity is of concern for future translational application. Alternatively, growth factors and hormones such as basic fibroblast growth factor (bFGF) and hydrocortisone have been reported to act as a therapeutic inducer for cardiomyocytes differentiation. In this study, we aim to investigate the ability of bFGF and hydrocortisone in combination to stimulate the differentiation of mesenchymal stem cells (MSC) into cardiomyocytes lineage. Sheep adipose tissue stem cell (ATSC) and bone marrow stem cell (BMSC) were isolated, cultured and induced with the three groups of induction factors; 5-aza alone, the combination of hydrocortisone and bFGF and all three factors in combination for cardiomyogenic differentiation. Morphological, protein and functional ability of both ATSC and BMSC were observed and analysed to confirm cardiomyocyte differentiation. Viability of BMSC and ATSC in each treated group was significantly higher (P < 0.05) on both cells after treated with 10 nM of bFGF and 50 μM of hydrocortisone. Cardiomyocyte proteins; α-Sarcomeric actin (αSA) and Phospolamban (Plb) was detected in both ATSC and BMSC exposed to induction factors but not in the control negative group. Both ATSC and BMSC without induction factors showed only minute cell number possesses αSA and Plb. Calcium ion (Ca²⁺) spark was observed in primary heart cells. Similarly, Ca²⁺ spark was also detected in induced ATSC and BMSC, proving some functionality of induced cells. In conclusion, bFGF and hydrocortisone are safer induction factor compared to the currently used 5-aza as both showed higher viability after induction, therefore more cells are available for future use in cardiac tissue engineering.

Keywords: 5-Azacytidine; basic Fibroblast Growth Factor; cardiomyocytes differentiation; hydrocortisone; stem cells

ABSTRAK

Sel induk boleh dibezakan menjadi kardiomiosit dengan aruhan 5-azasitidin (5-aza) tetapi sifat karsinogeniknya menimbulkan kerisauan bagi kegunaan klinikal pada masa hadapan. Sebagai alternatif, faktor pertumbuhan dan pelbagai jenis hormon seperti faktor pertumbuhan fibroblas asas (bFGF) dan hidrokortison dilaporkan boleh bertindak sebagai pemacu terapi untuk pembezaan kardiomiosit. Kajian ini bertujuan untuk mengkaji kemampuan bFGF dan hidrokortison secara gabungan untuk merangsang pembezaan MSC kepada leluhur kardiomiosit. Sel dasar lemak (ATSC) dan tulang sum-sum kambing (BMSC) diasingkan, dikultur dan diaruh dengan tiga kumpulan faktor aruhan; 5-aza sahaja, gabungan hidrokortison dan bFGF dan ketiga-tiga faktor gabungan untuk pembezaan kardiomogenik. Perubahan morfologi, protein dan fungsi kedua-dua ATSC dan BMSC dikaji dan dianalisis untuk mengesahkan pembezaan leluhur kardiomiosit. Perkembangan ATSC dan BMSC pada setiap kumpulan yang dirawat jauh lebih tinggi (P <0.05) pada kedua-dua sel setelah dirawat dengan 10 nM bFGF dan 50 μM hidrokortison. Protein kardiomiosit, α-Sarcomeric actin (αSA) dan Phospolamban (Plb) dikesan pada kedua-dua ATSC dan BMSC yang terdedah kepada faktor aruhan tetapi tidak dalam kawalan negatif. Kedua-dua sel, ATSC dan BMSC tanpa faktor aruhan menunjukkan hanya sebilangan kecil sel mempunyai αSA dan Plb. Percikan ion kalsium (Ca^{2 +}) diperhatikan pada sel jantung primer, yang turut dikesan pada ATSC dan BMSC yang diinduksi. Maka, sel yang diinduksi sedikit sebanyak berfungsi seperti kardiomiosit. Kesimpulannya, bFGF dan hidrokortison adalah faktor aruhan yang lebih selamat berbanding dengan 5-Aza yang digunakan sekarang. Hal ini demikian adalah kerana kedua-duanya menunjukkan perkembangan yang lebih tinggi selepas aruhan, oleh itu lebih banyak sel tersedia untuk kegunaan pada peringkat klinikal.

Kata kunci: 5-azasitidin; faktor tumbesaran fibroblas asas; hidrokortison; pembezaan kardiomiosit; sel dasar

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*Corresponding author; email: nadiahsulaiman@ukm.edu.my