Balb c nude mouse

All animal studies were conducted according to the protocols approved by the Institutional Animal Care and Use Committee of the Seoul National University. The human breast cancer cell line MDA-MB-231/HER2, a HER2-expressing variant of MDA-MB-231 (ATCC, Manassas, VA) was used for this research. Cancer cells were cultured with Dulbecco's modified Eagle's medium (DMEM; Welgene, Daegu, Korea) containing 10% Fetal Bovine Serum and 1% penicillin/streptomycin. Cells were incubated in a humidified atmosphere of 5% CO₂ at 37°C and passaged with 0.125% trypsin. Then, the MDA-MB-231/HER2 breast cancer cells were harvested, and 5 × 10⁶ cells were injected in the mammary fat pad of a female BALB/c nude mouse (OrientBio, Sungnam, Korea). A 30-gauge needle was used for injection. The injection volume was 0.1 mL containing tumor cells and medium/matrigel complex with 50% dilution. Tumor growth was evaluated for 14 days, and an in vivo tumor imaging was performed when the largest tumor diameter reached 1–1.5 cm.

All animal studies were approved by the Institutional Animal Care and Use Committee (IACUC) of The Catholic University of Korea (Approval number: CUMC-2015-0048-04). Myocardial infarction in the rodent model was induced as we previously described [28 (link),29 (link),30 (link)]. Male BALB/c nude mouse (8-week-old, 30–40 g; Orient bio, Seongnam, Korea) anesthetized via inhalation of 2% isoflurane were intubated through the trachea with an 18 G intravenous catheter. Then, the mice were mechanically ventilated with medical-grade oxygen. After the left thoracotomy was performed, MI was induced by the left anterior descending artery’s permanent ligation. Immediately after MI induction, BM-MSC cells, HGF-eMSC cells, or a mixture (BM-MSC and HGF-eMSC) of cells were injected directly into the ischemic area. To trace the BM-MSCs within the heart tissues further, the BM-MSCs were pre-labeled with a red fluorescence dye, CM-DiI, prior to cell injection. Briefly, BM-MSCs were stained using CellTracker™ CM-DiI Dye (Invitrogen™, Carlsbad, CA, USA). BM-MSCs were incubated in the CM-DiI Dye solution (5 µg/mL) for 5 min at 37 °C, followed by additional 15 min at 4 °C. After labeling, BM-MSCs were washed with phosphate-buffered saline (PBS) and re-suspend in serum free medium for further uses.

A BALB/c nude mouse (8 week-old female, Orientbio) was prepared for in vivo experiments. All animal experiments were carried out in accordance with the guidelines of Chonnam National University Hwasun Hospital. The mouse was anesthetized with a mixture of ketamine (87.5 mg/kg) and xylazine (12.5 mg/kg). B16F10 murine melanoma cells were harvested, pelleted by centrifugation at 300 g for 5 min, and suspended in PBS. The obtained melanoma cell suspension consisted of 1 × 10⁵ melanoma cells in 10 µL of PBS and was implanted subcutaneously into the right ear. Melanoma cells were injected into two regions, as shown in Figure 1a.