Sains Malaysiana 49(1)(2020): 113-119

http://dx.doi.org/10.17576/jsm-2020-4901-13

Photoreceptor Therapy: Generation of Neurosphere-Like Cells from Human Mesenchymal Stem Cells Expressing Erythropoietin

(Terapi Fotoreceptor: Generasi Sel Seperti Sfera Neuro daripada Sel Stem Mesenkima Manusia Ekspresi Eritropoletin)

MOK POOI LING^1,2,3*, SHIRLEY DING SUET LEE², AISHA FARHANA¹, BADR ALZAHRANI¹, MOHAMMED SAFWAN ALI KHAN² & SURESH KUMAR^3,4,5*

¹Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf University, Sakaka, P.O. Box 2014, Aljouf Province, Saudi Arabia

²Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

³Genetics and Regenerative Medicine Research Centre, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

⁴Department of Medical Microbiology and Parasitology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

⁵Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

Diserahkan: 6 September 2019/Diterima: 7 Oktober 2019

ABSTRACT

The loss of photoreceptors is a major concern implicated in age-macular degeneration (AMD), a type of neurodegenerative disorder. Failure to prescribe a suitable treatment due to the lack of understanding of the molecular pathogenesis, and limited capacity to compensate irreparably damaged photoreceptors in the retina have greatly contributed to the progression of visual dysfunction. Our previous study has shown that Mesenchymal Stem Cells (MSCs) expressing erythropoietin (EPO) could commit into photoreceptor cell lineage. However, the efficiency of cell differentiation is limited. The present study aims to explore the capacity of these MSCs to form neurospheres. The cells were transduced with lentiviral particles encoding for human EPO and green fluorescent protein (GFP) genes, culture-expanded and sorted before subjected for differentiation induction into neural precursor cells. Our results showed that MSC-EPO developed into larger neurosphere and expressed relatively higher expression of nestin compared with MSCs alone when cultured under neural induction medium. These preliminary findings suggested that MSC-EPO have greater neurogenic potential than MSCs alone. Further study is needed to evaluate the possibilities of neurosphere to differentiate into functional photoreceptor cells. We believe that the success of neurosphere expansion may potentially be useful in scaling up the manufacturing of photoreceptors in a shorter time and at an efficient cost for retinal cell replacement therapy.

Keywords: Erythropoietin; mesenchymal stem cells; neural differentiation; neurosphere; photoreceptor

ABSTRAK

Kehilangan fotoreseptor menjadi kebimbangan utama dalam degenerasi makro-usia (AMD), sejenis gangguan neurodegeneratif. Perkembangan disfungsi visual berlaku apabila terdapat kegagalan untuk memberi rawatan yang sesuai akibat kekurangan pemahaman dalam patogenesis molekul dan keupayaan terhad untuk mengganti kerosakan fotoreseptor di retina. Kajian terdahulu menunjukkan bahawa Sel Stem Mesenkima (MSC) yang mengungkapkan eritropoletin (EPO) boleh berkembang menjadi sel berketurunanan fotoreseptor. Walau bagaimanapun, kecekapan pembezaan sel adalah terhad. Kajian ini bertujuan untuk meneroka keupayaan MSCs untuk membentuk sfera neuro. Sel telah ditransduksi dengan zarah lentiviral yang mengekod gen manusia protein EPO dan hijau neon (GFP), diperbanyakkan melalui pengkulturan sel dan ditulen sebelum induksi diferensiasi ke sel prekursor saraf. Keputusan kami menunjukkan bahawa MSC-EPO berkembang menjadi sfera neuro yang lebih besar dan menunjukkan ekspresi nestin yang lebih tinggi berbanding dengan MSC sahaja apabila dikultur di dalam medium induksi saraf. Penemuan awal ini mencadangkan bahawa MSC-EPO mempunyai potensi neurogen yang lebih besar daripada MSC sahaja. Kajian lanjut diperlukan untuk menilai kebolehan sfera neuro untuk membeza seterusnya kepada sel-sel fotoreseptor yang berfungsi. Kami percaya bahawa kejayaan pengembangan sfera neuro berpotensi digunakan untuk meningkatkan produksi fotoreseptor dalam masa yang lebih singkat dan pada kos yang cekap untuk terapi penggantian sel retina.

Kata kunci: Eritropoletin; fotoreseptor; pembezaan neural; sel stem mesenkima; sfera neuro

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*Pengarang untuk surat-menyurat; email: rachelmok2005@gmail.com