Immunological properties of umbilical cord blood-derived mesenchymal stromal cells

https://doi.org/10.1016/j.cellimm.2008.04.003Get rights and content

Abstract

Mesenchymal stromal cells (MSCs) are promising candidates for developing cell therapies for intractable diseases. To assess the feasibility of transplantation with human umbilical cord blood (hUCB)-derived MSCs, we analyzed the ability of these cells to function as alloantigen-presenting cells (APC) in vitro. hUCB-MSCs were strongly positive for MSC-related antigens and stained positively for human leukocyte antigen (HLA)-AB and negatively for HLA-DR. When treated with interferon (IFN)-γ, the expression of HLA-AB and HLA-DR, but not the co-stimulatory molecules CD80 and CD86, was increased. hUCB-MSCs did not provoke allogeneic PBMC (peripheral blood mononuclear cell) proliferation, even when their HLA-molecule expression was up-regulated by IFN-γ pretreatment. When added to a mixed lymphocyte reaction (MLR), hUCB-MSCs actively suppressed the allogeneic proliferation of the responder lymphocytes. This suppressive effect was mediated by soluble factors. We conclude that hUCB-MSCs can suppress the allogeneic response of lymphocytes and may thus be useful in allogeneic cell therapies.

Introduction

MSCs are multipotent cells found in fetal liver, bone marrow (BM), and cord blood [1], [2], and have the capacity to differentiate in vitro into several mesodermal (bone, cartilage, tendon, muscle, and adipose), endodermal (hepatocyte), and ectodermal (neurons) tissues [3]. Classically, MSCs are defined as being able to adhere to plastic, expressing CD29, CD73, CD44, CD90, CD105, and major histocompatibility complex type [MHC] class I antigens, and not expressing the hematopoietic cell markers CD34, CD45, and MHC class II antigen. MSCs constitutively secrete a large number of cytokines and promote the expansion and differentiation of hematopoietic stem cells (HSCs) [4]. Furthermore, MSCs are not immunogenic, namely, they do not induce allogeneic lymphocytes to proliferate in vitro[5]. Indeed, MSCs appear to suppress these allogeneic proliferative responses. For example, in in vivo trials, co-infusion of MSCs as a third party delayed donor cell rejection, even when immunosuppressant drugs were not used [6]. These characteristics make MSCs potent candidates for the development of allogeneic cell-based therapeutic strategies. The therapeutic efficacy of MSCs for bone, joint, and neuronal diseases has recently been reported [7], [8].

BM is the main source of MSCs and BM-MSCs have already been used in various clinical studies. However, the number of MSCs in the BM and their multi-lineage differentiation capacity decline with age [9]. Therefore, human umbilical cord blood is often used as an alternative source of stem or progenitor cells, namely, HSCs and MSCs [10]. hUCBs also have a practical advantage over BM-derived MSCs in that they are obtained by non-invasive methods that do not harm either the mother or the infant [11]. Furthermore, cord blood stem cells are more immature than adult MSCs and expand readily in vitro[12]. Given these features along with their potent differentiation potential [3], hUCB-MSCs are an attractive source for cellular or gene transfer therapy. Moreover, a female patient with chronic spinal cord injury who was given hUCB-MSCs showed functional and morphological improvement [13]. The transfer of hUCB-MSCs was also useful in treating Buerger’s disease [14].

The aim of the present study was to investigate the immunological properties of hUCB-MSCs to assess their potential usefulness in allogeneic transplantation. We examined the effect of IFN-γ pretreatment on the expression of MHC molecules on the hUCB-MSC cell surface and assessed whether hUCB-MSCs can provoke an in vitro allogeneic reaction. We also asked whether hUCB-MSCs, like other MSCs [15], [16], can inhibit allogeneic MLR and mitogen-induced lymphocyte proliferation. In addition, we examined whether these immunological properties of hUCB-MSCs change after they differentiate into various lineages.

Section snippets

Isolation and culture of hUCB-MSCs

This study was approved by the Institutional Review Board of Medipost Inc., Seoul, Korea. The hUCB-MSCs were separated and maintained as described previously [3]. Briefly, hUCB samples were collected from the umbilical vein of deliveries with informed maternal consent. Mononuclear cells were isolated from the hUCBs by centrifugation through a Ficoll-Hypaque gradient (density 1.077 g/cm3, Sigma, St. Louis, MO, USA). The separated mononuclear cells were washed, suspended in α-minimum essential

Characterization of the hUCB-MSCs

MSCs were successfully isolated from three hUCBs and could be separately cultured until passage 4. As reported previously [3], the hUCB-MSCs had a fibroblast-like morphology (Fig. 1A). Three different hUCB-MSC clones were uniformly positive for MSC-related antigens, namely, CD73 (96.1 ± 2.3%), CD105 (91.3 ± 4.1%), CD90 (96.8 ± 1.5%), CD166 (93.1 ± 3.5%), CD29 (91.1 ± 6.7%), and CD44 (92.1 ± 5.1%) (Fig. 1D). In addition, these cells were positive for HLA-class I (HLA-AB; 98.1 ± 3.3) but expressed neither

Discussion

hUCB is currently used as an alternative to BM as a source of stem cells for management of hematological malignancies. The possible usefulness of hUCB-MSCs in many preclinical models ranging from myocardial ischemia and stroke to cartilage regeneration is also under intense experimental investigation [7], [18]. The main theoretical limitation hampering the clinical transplantation of hUCB cells would be graft rejection due to the immunological barrier imposed by HLA mismatching. However, in

Acknowledgments

This research was supported by a Grant (SC3190) from the Stem Cell Research Center of the 21st Century Frontier Research Program funded by the Ministry of Science and Technology, Republic of Korea.

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    These authors contributed equally to this work.

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