A Simple Method to Isolate and Expand Human Umbilical Cord Derived Mesenchymal Stem Cells: Using Explant Method and Umbilical Cord Blood Serum

BACKGROUND AND OBJECTIVES: Mesenchymal stem cells (MSCs) are multipotent stem cells that can be isolated from umbilical cords and are therapeutically used because of their ability to differentiate into various types of cells, in addition to their immunosuppressive and anti-inflammatory properties. Fetal bovine serum (FBS), considered as the standard additive when isolating and culturing MSCs, has a major limitation related to its animal origin. Here, we employed a simple and economically efficient protocol to isolate MSCs from human umbilical cord tissues without using digestive enzymes and replacing FBS with umbilical cord blood serum (CBS). METHODS AND RESULTS: MSCs were isolated by culturing umbilical cord pieces in CBS or FBS supplemented media. Expansion and proliferation kinetics of cells isolated by explant method in the presence of either FBS or CBS were measured, with morphology and multi-differentiation potential of expanded cells characterized by flow cytometry, RT-PCR, and immunofluorescence. MSCs maintained morphology, immunophenotyping, multi-differentiation potential, and self-renewal ability, with better proliferation rates for cells cultured in CBS compared to FBS supplement media. CONCLUSIONS: We here present a simple, reliable and efficient method to isolate MSCs from umbilical cord tissues, where cells maintained proliferation, differentiation potential and immunophenotyping properties and could be efficiently expanded for clinical applications.

The stimulative effects of cord blood serum on hematopoietic cells in vitro / 中国免疫学杂志

Objective:To study the effects of human CBS(Cord Blood Serum) on hematopoietic cells culture in vitro.Methods:CBS and/or different combinations of cytokines were added to the culture system derived from human bone marrow mononuclear cells.Colony forming capacity of colony forming unit-granulocyte/ monocyte (CFU-GM)?colony forming unit-granulocyte/ erythrocyte/ megakaryocyte/ monocyte (CFU-GEMM)?cobble-stone area forming cells (CAFC)?long-term culture initiating cells (LTC-IC) were observed.Results:CBS had certain cytokine-like stimulating activity that could promote the proliferation and differentiation of hematopoietic cells of bone marrow in vitro. By comparison with cytokines, its GM-CSF-like activity was equal to 65.6 ?g/L rhGM-CSF, and its IL-3-like activity was equal to 2.9 ?g/L rhIL-3.Conclusion:CBS contains stimulating activities for proliferating and differentiating of hematopoietic cells. CBS alone can maintain hematopoietic cells culture in vitro. This study showed the prospect of clinical application of CBS.

THE RELATIONSHIP OF SERUM CALCIUM, MAGNESIUM, ZINC, AND COPPER BETWEEN MATERNAL AND CORD BLOOD / 营养学报

Serum calcium, magnesium, zinc, and copper were determined in 117 healthy pregnant women and their infants. Maternal and cord blood were collected at delivery and assayed by flame atomic absorption spectropho-tometry, and the data obtained were evaluated by regression analysis.The results suggested that the serum concentrations of calcium, magnesium, and zinc of maternal blood were significantly lower than that of cord blood. The concentrations of serum calcium and magnesium between maternal and cord blood were highly correlated, but not of serum zinc. The serum concentration of copper in maternal blood was significantly higher than that in cord blood, but there was nos ignificant correlation between them. The serum calcium of infants always maintained over 2.0mmol/L(8mg/dl), even when the serum calcium of pregnant women was lower than 2.0mmol/L (8mg/dl). This fact indicates that the fetus draw much calcium from mother for development, despite calcium in the mother being depleted.