Optimization of human umbilical cord blood-derived mesenchymal stem cell isolation and culture methods in serum- and xeno-free conditions

Background Although umbilical cord blood (UCB) is identified as a source of mesenchymal stem cells (MSCs) with various advantages, the success in cell isolation is volatile. Therefore, it is necessary to optimize methods of cord blood-derived MSC (UCB-MSC) isolation and culture. In this study, we evaluated the efficiency of UCB-MSC isolation and expansion using different commercially available serum- and xeno-free media and investigated the capacity of autologous serum and plasma as a supplement to support cell proliferation. Additionally, we defined the presence of multilineage-differentiating stress-enduring (Muse) cells in the UCB-MSC population. Functions of UCB-MSC in in vitro angiogenesis processes and anti-cancer were also verified. Methods Mononuclear cells were isolated using density gradient separation and cultured in four commercial media kits, as well as four surface coating solutions. UCB-MSCs were characterized and tested on tube formation assay, and co-cultured with SK-MEL cells in a transwell system. Results The results showed that only StemMACS™ MSC Expansion Media is more appropriate to isolate and culture UCB-MSCs. The cells exhibited a high cell proliferation rate, CFU forming capability, MSC surface marker expression, trilineage differentiate potential, and chromosome stability. In addition, the culture conditions with autologous serum coating and autologous plasma supplement enhanced cell growth and colony forming. This cell population contained Muse cells at rate of 0.3%. Moreover, UCB-MSCs could induce the tube formation of human umbilical vein endothelial cells and inhibit more than 50% of SK-MEL cell growth. Conclusions UCB-MSCs could be high-yield isolated and expanded under serum- and xeno-free conditions by using the StemMACS™ MSC Expansion Media kit. Autologous serum coating and plasma supplement enhanced cell proliferation. These UCB-MSCs had effected the tube formation process and an anti-cancer impact.

Comparative assessment of umbilical cord blood with peripheral venous blood using hematological scoring system as an early predictive screening method for the detection of early-onset neonatal sepsis in the tertiary care center of Central India

Background: Sepsis is one of the major causes of neonatal morbidity and mortality. Early recognition and diagnosis of early-onset neonatal sepsis (EONS) is difficult because of the variable and non-specific clinical presentation of this condition. Hence, there is a need for early predictive screening method for EONS. Aims and Objectives: To compare the umbilical cord blood Haematological Scoring System (HSS) with peripheral venous blood as an early predictive screening method for detection of EONS. Materials and Methods: 100 inborn neonates with two or more risk factors for EONS, chosen by sequential sampling method were included in this prospective analytical study. Blood samples were collected from the umbilical cord and peripheral vein analyzed for hematological parameters, sepsis screen, and peripheral smear for HSS of Rodwell et al., send for blood culture. Blood cultures were performed as gold standard for diagnosing neonatal sepsis and sepsis screen was done to corroborate the diagnosis of neonatal sepsis. Results: Of 100 neonates, 21 belongs to sepsis; 14 to probable sepsis; 65 to no sepsis. HSS in umbilical cord blood (UCB) had Sensitivity-74.28%, Specificity-92.30%, PPV-83.87%, NPV-86.95% and HSS in PVB had Sensitivity-62.85%, Specificity- 87.69%, PPV-75.86%, NPV-81.69%. Conclusion: HSS score in UCB can be used as a simple, quick, cost-effective, and readily available screening test with decent sensitivity and high specificity, for the detection of EONS.