Poietics human mscs

Poietics human MSCs (passage 2) were purchased from Lonza (Basel, Switzerland), expanded until passage 4, and used as normoxic MSCs (norMSCs). The cells were cultured and passaged with StemPro MSC SFM XenoFree (Thermo Fisher Scientific, Waltham, MA, USA) under normal oxygen conditions (21% O₂) and 5% CO₂ at 37 °C. Passage 4 cells were frozen and stored using liquid nitrogen. StemPro BM MSCs (passage 4; Thermo Fisher Scientific) were purchased from Thermo Fisher Scientific and used as hypoxic MSCs (hypoMSCs). The cells were cultured in StemPro MSC SFM XenoFree (Thermo Fisher Scientific) under hypoxic conditions (5% O₂) and 5% CO₂ at 37 °C. Cryopreservation solution with 10% DMSO was used for cryopreservation of these cells.

Primary human mesenchymal cells (Poietics™ human MSCs, Lonza, Wlakersville. Inc.) were obtained from a commercial source and grown using the proprietary protocol provided by Lonza. Mesenchymal stem cells were cultured at cell seed densities of 2 × 10⁶ – 4 × 10⁶/cm² in Dulbecco’s Modified Eagle Medium (DMEM (1X) + GlutaMAX; Gibco; Life Technologies) with 10% FBS and supplemented with penicillin and streptomycin. The growth of hMSCs for seven days preceded encapsulation. A single culture dish of hMSCs after seven days of culture contained 2 million cells at an approximately 95% confluence level. hMSCs were trypsinized and pelleted before the addition of 1 ml of a 5% alginate solution.