case_images: PMC8175829_01
case_images: Study design. (A) Umbilical cord-derived mesenchymal stromal cells (UC-MSC) were isolated, expanded, and evaluated for surface markers, viability, presence of fungi and bacteria, and chromosomal abnormalities. The patient received three infusions of 500.00 UC-MSCs/kg and two infusions of 400 mg of tocilizumab. The patient's clinical and laboratory evaluations were performed pre-MSC infusion (D1) and on days 2, 4, 6, 14, 60, and 120. (B) Schematic representation of the days before (minus) and after the first infusion of UC-MSC.
case_images: 10.1177_09636897211021008-fig1.jpg
case_images: PMC8175829_01_10.1177_09636897211021008-fig1.jpg
case_images: fig1-09636897211021008
case_images: Ten days after the beginning of symptoms and three days of the worsening of the severity of the clinical condition (D0), the patient was included in a study protocol for treatment with allogeneic umbilical cord-derived mesenchymal stromal cells (UC-MSC) to received three intravenously infusions of 500,000 cells (passage 3) per kilogram in alternate days (Fig. 1A, B).
case_images: Quality control of umbilical cord-derived mesenchymal stromal cells (UC-MSC). (A) Cell characterization; (B) Cell viability. (C) Cytogenetics test. The cells showed a high expression of CD29, CD73, CD90, and low expression of CD14, CD19, CD34, CD45, and HLA-DR markers (A). Cell viability analysis showed less than 8% of dead cells (7-AAD) and less than 1% of cells in apoptosis (Annexin-V) (B). The UC-MSC did not show any chromosomal aberrations. Ann: Annexin; 7-AAD: 7-amino actinomycin.
case_images: 10.1177_09636897211021008-fig2.jpg
case_images: PMC8175829_01_10.1177_09636897211021008-fig2.jpg
case_images: fig2-09636897211021008
case_images: Advanced Cell Therapy Product. After the standardized process of mechanical and enzymatic isolation, MSC were expanded in vitro until passage 3 and showed the viability of 97%; negative cultures for bacteria and fungi, absence of endotoxin and Mycoplasma; flow cytometry characterization with positive markings for CD90, CD105, CD29, and CD73 and negative for CD45, CD34, CD14, CD19, and HLA-DR (BD Biosciences, San Jose, CA, USA); and absence of clonal chromosomal aberrations in G-banding karyotype analysis (Fig. 2).