The Role of CCL5 in the Ability of Adipose Tissue-Derived Mesenchymal Stem Cells to Support Repair of Ischemic Regions

Abstract

Mesenchymal stem cells (MSC) are multipotent and possess high proliferative activity, and thus are thought to be a reliable cell source for cell therapies. Here, we isolated MSC from adult tissues-bone marrow (BM-MSC), dental tissue (DT-MSC), and adipose tissue (AT-MSC)-to compare how autotransplantation of these MSC effectively supports the repair of bone fracture and ischemic tissue. An analysis by in vitro differentiation assays showed no significant difference among these MSC. The degree of calcification at the joint region of bone fracture was higher in mice transplanted with AT-MSC than in mice transplanted with BM-MSC or DT-MSC. To compare the abilities of MSC, characterize how those MSC affect the repair of ischemic tissue, vascular occlusion was performed by ligation of the femoral artery and vein. Of note, the blood flow in the ischemic region rapidly increased in mice injected with AT-MSC, as contrasted with mice injected with BM-or DT-MSC. The number of CD45-and F4/80-positive cells at the femoral region was higher in AT-MSC recipients than in recipients of BM-MSC or DT-MSC. We evaluated the mRNA expression of angiogenic and migration factors in MSC and found the expression of CCL5 mRNA was higher in AT-MSC than in BM-MSC or DT-MSC. Transplantation of AT-MSC with impaired expression of CCL5 clearly showed a significant delay in the recovery of blood flow compared with the control. These findings have fundamental implications for the modulation of AT-MSC in the repair of vasculature and bone fracture.

Increased Expression of EGR-1 in Diabetic Human Adipose Tissue-Derived Mesenchymal Stem Cells Reduces Their Wound Healing Capacity

“…Human adipose tissue was obtained after obtaining informed consent from diabetic (n = 3, HbA1c >7.0, longterm treatment) and nondiabetic (n = 3) donors, male, age = 59 -10 years, who were undergoing procedures in the Department of Cardiovascular Surgery, University of Tsukuba Hospital, Tsukuba, Japan. The isolation of AT-MSCs was performed as previously described [24]. Cells were cultured in Iscove's modified Dulbecco's medium (IMDM, 12200-069; Invitrogen), 10% fetal bovine serum, 2 mg/mL Lglutamine (25030; Invitrogen), and 5 ng/mL recombinant human bFGF (064-04541; PeproTech) at 37°C in 5% CO 2 and a humidified atmosphere.…”

“…In vitro differentiation was performed as described previously [24]. Osteogenic and adipogenic differentiation were assessed on day 21 by 1% Alizarin Red S (Kodak) and Oil Red O (Muto Pure Chemicals) staining, respectively.…”

A Chemokine Receptor, CXCR4, Which Is Regulated by Hypoxia-Inducible Factor 2α, Is Crucial for Functional Endothelial Progenitor Cells Migration to Ischemic Tissue and Wound Repair

“…We previously found that Alde-Low EPCs were incorporated into newly formed neovessels, indicating that transplanted EPCs contribute to the revascularization of ischemic tissue at the site of the skin flap rather than through the promotion of neoangiogenesis. In the case of mesenchymal stem cells (MSCs), we reported that, in addition to Alde-Low EPCs, transplanted adipose tissue-derived MSCs (AT-MSCs) effectively repaired the perfusion of the blood flow in a mouse model of hindlimb ischemia, indicating that AT-MSC promoted neoangiogenesis through a paracrine mechanism [ 25 ]. The migrated EPCs can secrete SDF-1 themselves to recruit the additional migration of CXCR4-positive EPCs or inflammatory cells to the site of ischemic tissues, suggesting that transplanted and migrated EPCs may function through a paracrine mechanism.…”

Rejuvenation of mesenchymal stem cells by extracellular vesicles inhibits the elevation of reactive oxygen species

“…Human adipose tissues were collected from the infants (n = 5, 1-12 months) and elderly donors (n = 5, 70-80 years old) who are undergoing the treatment in Department of Cardiovascular Surgery, University of Tsukuba Hospital, Tsukuba, Japan. AT-MSCs were isolated according to a previously described method 44 . In brief, adipose tissues were minced and digested with 0.1% collagenase (Invitrogen, Waltham, Massachusetts, CA, USA) in phosphate-buffered saline (PBS) at 37 °C for 1 h. AT-MSCs were harvested by the centrifugation at 1600 rpm in 7 min, then suspended in the culture medium (Iscove's Modified Dulbecco Medium [IMDM]; Invitrogen), containing 10% heat-inactivated fetal bovine serum (FBS; Invitrogen), 2 mg/ml L-glutamine (Invitrogen), 100 units/ml penicillin (Invitrogen), and 5 ng/ml bFGF (Peprotech, Cranbury, NJ, USA).…”

“…Total RNA from AT-MSCs was isolated using Sepasol-RNA I Super G (Nacalai Testque, Kyoto, Japan) according to the previous described method 44 . In order to synthesize cDNA, a RT-PCR kit (Toyobo Co., Ltd, Osaka, Japan) was used with 1 µg of total RNA.…”