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
Diserahkan: 25 Februari
2020/Diterima: 9 Oktober 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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*Pengarang untuk surat-menyurat;
email: angela@ppukm.ukm.edu.my
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