Sains Malaysiana 50(6)(2021): 1715-1726

http://doi.org/10.17576/jsm-2021-5006-18

Immunomodulatory Properties of Wharton’s Jelly-Derived Mesenchymal Stem Cells from Three Anatomical Segments of Umbilical Cord

(Sifat Imunomodulasi Sel Stem Mesenkima Jeli Wharton yang Dipencilkan daripada Tiga Segmen Anatomi Tali Pusat)

JEZAMINE LIM¹, SUE PING ENG¹, WEI YEN YEOH¹, YIK WAN LOW¹, NUR MOHD SHAFWAN BIN JUSOH¹, AIN SYAHIRAH BINTI RAHMAT¹, AMIRAH SHAHRANI¹, FAIQ BAHRANI YAHYA¹, RUSHDA ADIBA ABDUL RAHMAN¹, ZAINUL RASHID MOHAMAD RAZI², CHOOI FUN LEONG³, SHINSMON JOSE¹ & MIN HWEI NG¹*

¹Centre for Tissue Engineering & Regenerative Medicine, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, 56000 Kuala Lumpur, Federal Territory, Malaysia

²Department of Obstetrics and Gynaecology, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, 56000 Kuala Lumpur, Federal Territory, Malaysia

³Department of Haematology, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, 56000 Kuala Lumpur, Federal Territory, Malaysia

Received: 25 February 2020/Accepted: 9 October 2020

ABSTRACT

Mesenchymal stem cells (MSCs) are multipotent progenitor cells that are reported to be immune-privileged and immune-evasive. MSCs are capable of differentiating into specific cell types for subsequent use in cell-based therapy. They express low levels of human leucocyte antigen (HLA)-ABC and no HLA-DR. Wharton’s jelly-derived MSCs (WJ-MSCs) were also found to express human leukocyte antigen G (HLA-G), which renders them immunosuppressive. This study aimed to determine whether cultured WJ-MSCs retain their immune-privileged and immune-evasive properties after cell differentiation, and whether these properties differ among MSCs derived from different anatomical segments of the umbilical cord. Umbilical cords of healthy pregnant mothers undergoing caesarean section were obtained and grouped by three anatomical segments: fetal, middle, and maternal segments. WJ-MSCs were isolated, culture-expanded, and differentiated into osteogenic cells. Expression of HLA-DR, HLA-ABC, and HLA-G were quantified using flow cytometry. Both undifferentiated and osteodifferentiated WJ-MSCs were subsequently co-cultured with allogeneic peripheral blood mononuclear cells with/without lipopolysaccharide (LPS) stimulation for five days. Lymphocyte proliferation assay was performed using carboxyfluorescein succinimidyl ester (CFSE) as a tracker. Our results showed no significant difference existed in the HLA profiles among WJ-MSCs from different segments and between WJ-MSCs with and without osteogenic differentiation. Mean levels for HLA-G, HLABC, and HLA-DR were 24.82±17.64, 52.50±18.41, and 1.00±1.68%, respectively. Stimulation with LPS and WJ-MSCs increased peripheral blooc mononuclear cells (PBMC) proliferation. However, PBMC proliferation was significantly lower when PBMCs were co-cultured with osteodifferentiated WJ-MSCs (p < .05; with LPS stimulation and p < .001 without LPS stimulation) than when they were co-cultured with undifferentiated WJ-MSCs. These findings suggest that cultured WJ-MSCs stimulate lymphocyte proliferation and are not immune-privileged. Osteodifferentiated WJ-MSCs reduced the immunogenicity of WJ-MSCs, and this reduction in PBMC proliferation was even more pronounced in the presence of LPS (p < .05). In conclusion, cultured WJ-MSCs are not immune-privileged. Osteodifferentiated WJ-MSCs are less immunogenic than undifferentiated WJ-MSCs, in which case hypoimmunogenicity is more profound under LPS-stimulated conditions.

Keywords: Immunomodulation; peripheral blood mononuclear cells; umbilical cord; Wharton’s jelly mesenchymal stem cell

ABSTRAK

Sel Stem Mesenkima (MSCs) adalah sel progenitor multipoten yang boleh membeza menjadi sel yang khusus dan ini berpotensi digunakan untuk membaikpulih tisu yang rosak. MSC juga mempunyai sifat menindas atau mengelak gerak balas imun kerana ia mengungkap antigen leukosit manusia kelas I iaitu HLA-ABC yang rendah malah tidak mengungkap antigen leukosit manusia kelas II iaitu HLA-DR. Satu keistimewaan sel stem mesenkima yang dipencil daripada bahagian jeli Wharton tali pusat (WJ-MSCs) ialah ia juga menggunakan HLA-G, sejenis antigen leukosit kelas I dan ini menyebabkan WJ-MSC mempunyai daya menindas gerak balas imun yang lebih tinggi. Dalam kajian ini, matlamat kami adalah untuk menentukan sama ada WJ-MSC yang dipencilkan daripada segmen tali pusat yang berlainan dapat mengekal keistimewaan keimunan asalnya. Tali pusat daripada ibu yang sihat dan menjalani pembedahan caesarean telah diperoleh dan dibahagikan kepada tiga segmen anatomi, iaitu segmen bahagian fetus, tengah dan ibu. Selepas itu, WJ-MSCs dipencilkan, dikultur dan dibezakan kepada sel osteogenik. Ekspresi HLA-DR, HLA-ABC dan HLA-G diukur melalui sitometri aliran. WJ-MSCs yang dibezakan kepada sel osteogenik atau MSC asli dibiakkan bersama dengan sel mononuklear darah alogenik dengan atau tanpa rangsangan lipopolisakarida (LPS) selama 5 hari. Ujian proliferasi sel limfosit dijalankan menggunakan ester karboksifluoresein sinksinmidil (CFSE) sebagai penanda jejak. Hasil daripada kajian ini menunjukkan bahawa tiada perbezaan yang signifikan dalam profil HLA antara WJ-MSCs daripada segmen yang berbeza dan juga antara WJ-MSC asli dan WJMSC pembezaan osteogenik. Purata ungkapan HLA-G, HL-ABC dan HLA-DR masing-masing adalah 24.82.0±17.64, 52.50±18.41, 1.00±1.68%. Penambahan LPS merangsang proliferasi sel limfosit. Tetapi, apabila sel limfosit dikultur bersama dengan WJ-MSC daripada segmen yang berbeza, proliferasi sel limfosit menurun (p < .05; dengan rangsangan LPS) dan (p < .001 tanpa rangsangan LPS). Penemuan ini mencadangkan bahawa WJ-MSC sebenarnya tidak dapat mengelak gerak balas imun. WJ-MSC pembezaan osteogenik dapat mengurangkan keimunogenan WJ-MSC dan penurunan proliferasi sel limfosit adalah lebih ketara apabila sel limfosit dirangsang oleh LPS (p < .05). Secara kesimpulan, WJ-MSC tidak dapat mengelak gerak balas imun. WJ-MSC pembezaan osteogenik kurang keimunogenan daripada WJ-MSC asli, lebih-lebih lagi jika dirangsang oleh LPS.

Kata kunci: Keimunogenan; sel limfosit; sel stem mesenkima jeli Wharton; tali pusat

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*Corresponding author; email: angela@ppukm.ukm.edu.my