Standardizing Umbilical Cord Mesenchymal Stromal Cells for Translation to Clinical Use: Selection of GMP‐Compliant Medium and a Simplified Isolation Method

Abstract

Umbilical cord derived mesenchymal stromal cells (UC-MSCs) are a focus for clinical translation but standardized methods for isolation and expansion are lacking. Previously we published isolation and expansion methods for UC-MSCs which presented challenges when considering good manufacturing practices (GMP) for clinical translation. Here, a new and more standardized method for isolation and expansion of UC-MSCs is described. The new method eliminates dissection of blood vessels and uses a closed-vessel dissociation following enzymatic digestion which reduces contamination risk and manipulation time. The new method produced >10 times more cells per cm of UC than our previous method. When biographical variables were compared, more UC-MSCs per gram were isolated after vaginal birth compared to Caesarian-section births, an unexpected result. UC-MSCs were expanded in medium enriched with 2%, 5%, or 10% pooled human platelet lysate (HPL) eliminating the xenogeneic serum components. When the HPL concentrations were compared, media supplemented with 10% HPL had the highest growth rate, smallest cells, and the most viable cells at passage. UC-MSCs grown in 10% HPL had surface marker expression typical of MSCs, high colony forming efficiency, and could undergo trilineage differentiation. The new protocol standardizes manufacturing of UC-MSCs and enables clinical translation.

Impact of the type of microcarrier and agitation modes on the expansion performances of mesenchymal stem cells derived from umbilical cord

“…Moreover, considering that WJ‐MSC have the age of the newborn, less impact due to the donor age is expected. Until now, WJ‐MSC have been successfully isolated and cultivated, in culture scales up to 800 ml …”

Characterization of Human AB Serum for Mesenchymal Stromal Cell Expansion

“…PL thus far can be considered the most used supplement for GMP-compliant MSC expansion instead of FBS. Several studies have demonstrated the efficacy of PL in supporting MSC cell growth [21,[30][31][32][33][34]. Heathman and colleagues [34] have shown that PL can be even better than FBS, delivering an increased cell growth in monolayer and also in microcarrier culture.…”

Therapeutic potential of mesenchymal stem cells for diabetes

“…Thus far, no standardized method for the isolation, characterization, expansion, potency testing, nor pathogen screening for MSCs exists (Arutyunyan et al 2016; Smith et al 2016; Weiss et al 2016). The regulation of cell based products by the US Food and Drug Administration (FDA) focuses on three main themes: i) prevention of transmitting communicable disease via contaminated tissue, ii) proper handling and processing of tissue, and iii) demonstration of clinical safety and effectiveness of cells, especially after extensive manipulation.…”

“…The FDA also requires tissue processing facilities to register, list their products, and provide accurate labeling of the products. Recent review articles have presented specifics focusing on standardization and production of clinical-grade stem cells (Giancola et al 2012; Sensebé et al 2013; Arutyunyan et al 2016; Smith et al 2016; Weiss et al 2016). …”