title: 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.
authors.affiliation: The Midwest Institute for Comparative Stem Cell Biotechnology, Department of Anatomy and Physiology, Kansas State University, Manhattan, KS 66506, USA.
authors.forename: J Robert
authors.initials: JR
authors.lastname: Smith
authors.affiliation: The Midwest Institute for Comparative Stem Cell Biotechnology, Department of Anatomy and Physiology, Kansas State University, Manhattan, KS 66506, USA.
authors.forename: Kyle
authors.initials: K
authors.lastname: Pfeifer
authors.affiliation: The Midwest Institute for Comparative Stem Cell Biotechnology, Department of Anatomy and Physiology, Kansas State University, Manhattan, KS 66506, USA; Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, 35390 Giessen, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Petry
authors.affiliation: The Midwest Institute for Comparative Stem Cell Biotechnology, Department of Anatomy and Physiology, Kansas State University, Manhattan, KS 66506, USA.
authors.forename: Natalie
authors.initials: N
authors.lastname: Powell
authors.affiliation: The Midwest Institute for Comparative Stem Cell Biotechnology, Department of Anatomy and Physiology, Kansas State University, Manhattan, KS 66506, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Delzeit
authors.affiliation: The Midwest Institute for Comparative Stem Cell Biotechnology, Department of Anatomy and Physiology, Kansas State University, Manhattan, KS 66506, USA.
authors.forename: Mark L
authors.initials: ML
authors.lastname: Weiss