Characterization of Fetal Tissue-derived Mesenchymal Stem Cells

Abstract

Mesenchymal stem cells (MSCs) have unique immunologic properties that may someday prove useful in cell-based therapy for various degenerative diseases. Its potential is limited, however, by several factors, including the rarity of these cells and difficulty in isolating them. To evaluate their potential as new sources for cell therapy, we isolated MSCs from human fetal tissue (hfMSC) derived from spontaneous abortus (8∼10 weeks) then studied their cell cycle and cell surface marker expression using a fluorescence-activated cell sorter (FACS), as well as the expression of differentiation markers using real-time polymerase chain reaction (RT-PCR). The hfMSCs were able to undergo PCR up to 20 times without displaying significant changes in morphology or expression of various stemness markers (Nanog and human telomerase reverse transcriptase [hAFP]), including germ layer markers (hNF68, alpha-cardiac actin, and hAFP). Also, teratomas were not seen in mice with severe combined immunodeficiency syndrome (SCID) that received a transplantation of hfMSCs with hTERT activity. The FACS analysis revealed that the majority of hfMSCs express mesenchymal markers CD13, CD44, CD71, CD90, CD105, CD253a, and HLA-ABC, but did not express CD31, CD34, CD38, CD45, and HLA-DR. Interestingly, hfMSCs derived from the cell membrane during early passages were negative for both HLA-ABC and HLA-DR, although HLA-ABC expression was detected during later passages (＞20 passages). We found that hfMSCs could be differentiated into an osteogenic lineage; this was indicated by modulation of osteoblast markers specific for mRNA. We conclude that hfMSCs could be used as a new source of cells to treat patients with osteogenic diseases, as well as to understand the mechanisms of immunosuppression by MSCs.

Mesenchymal stem cells: Cell therapy and regeneration potential

“…Unlike adult MSCs (i.e., BM‐MSCs), perinatal MSCs do not display major histocompatibility complex class II antigens (Anastasiu et al, ; Fariha, Chua, Tan, Tan, & Hayati, ; J. Kim, Jo, Kim, & Hwang, ; S. Liu, Yuan, et al, ; Maleki, Ghanbarvand, Reza Behvarz, Ejtemaei, & Ghadirkhomi, ; K.‐S. Shin et al, ; Tancharoen et al, ), making them more suitable for cell therapy.…”

“…Even the MSCs isolated from UC and FT have been reported to vary in their differentiation into various lineages (Beeravolu et al, ; Beeravolu et al, ; Brady et al, ; Guillot, Gotherstrom, Chan, Kurata, & Fisk, ; K.‐S. Shin et al, ). Controversy regarding the differentiation potential of MSCs may be due to the variation in the niche and unreliable in vitro techniques used to differentiate and characterize MSCs.…”

“…This review focuses on the current perspective, opportunities, and potential challenges for the use of MSCs for cell therapy and regenerative medicine applications. (Batsali et al, 2017;Beeravolu et al, 2017;Im et al, 2005;Jin et al, 2013;Vishnubalaji et al, 2012) PB Up to P5 27 hr 50% (Jumi Kim, Shin, Jeon, Chung, & Chae, 2012) UCB Up to P18 40 hr 24% (Heo, Choi, Kim, & Kim, 2016;Jin et al, 2013;Pievani et al, 2014) CL Up to P22 54 hr 59% (Araujo et al, 2017;Beeravolu et al, 2016;Beeravolu et al, 2017) WJ Up to P20 43 hr 80% (Batsali et al, 2017;Beeravolu et al, 2016;Beeravolu et al, 2017;Trivanovic et al, 2013) CPJ > P40 16 hr 92% (Bakshi, McKee, Walker, Brown, & Chaudhry, 2018;Beeravolu et al, 2016;Beeravolu et al, 2017) PC Up to P10 51 hr 16% (Araujo et al, 2017;Beeravolu et al, 2016;Beeravolu et al, 2017;Heo et al, 2016) FT-blood Up to P20 50 hr ND (Campagnoli et al, 2001;In't Anker et al, 2004) FT-BM Up to P20 33 hr 34% (Brady et al, 2014;Campagnoli et al, 2001;K.-S. Shin et al, 2009;Q. Wang et al, 2016) FT-heart Up to P15 80 hr ND (Garikipati et al, 2018) FT-liver Up to P20 52 hr ND (Campagnoli et al, 2001;In't Anker et al, 2004) FT-pancreas Up to P30 30 hr ND (Hu et al, 2003) AF Up to P7 39 hr 15% (Moraghebi et al, 2017) Abbreviations: AF, amniotic fluid; AT, adipose tissue; BM, bone marrow; CL, cord lining; CPJ, cord placenta junction; FT, fetal tissue; MSCs, mesenchymal stromal/stem cells; ND; not determined; P, passage; PB, peripheral blood; PC, placenta; UCB, umbilical cord blood; WJ, Wharton's jelly 2 | ADULT MSCs Bone marrow (BM) MSCs were first disc...…”

The transplantation of mesenchymal stem cells derived from unconventional sources: an innovative approach to multiple sclerosis therapy

“…Compared to BMMSCs or AD-MSCs, fetal tissue was found to contain a sufficient number of stem cells and progenitor cells for the development and a faster rate of proliferation (Deuse et al 2011;Ishii and Eto 2014). Moreover, FA-MSCs exhibit rare chromosomal changes, a stable karyotype, late senescence, and incapacity to generate teratomas that suggest their genetic stability (Shin et al 2009). In addition, these cells are poorly antigenic, expressing HLA-G for immune tolerance during pregnancy and thus are less likely to be rejected by transplant receivers (Hunt et al 2005).…”

“…Results of flow cytometry revealed that cells isolated from human umbilical cord, amniotic fluid, and placenta expressed CD13, CD29, CD44, CD73, CD90, CD105, and HLA-ABC, but not hematopoietic or endothelial specific antigens CD14, CD31, CD34, CD38, CD45, CD106, CD133, and HLA-DR (De Coppi et al 2007;Shin et al 2009;Christodoulou et al 2013) (Table 1). It is commonly accepted that potency of isolated FA-MSCs is related to the content of secretome, which includes G-CSF, TGF-β, VEGF, IGF1, BDNF, and GDNF.…”