Evaluation of reagents used to coat the hollow-fiber bioreactor membrane of the Quantum® Cell Expansion System for the culture of human mesenchymal stem cells

Abstract

The addition of a coating reagent to promote cell adherence is necessary to prepare the membrane surface of the Quantum® Cell Expansion System hollow-fiber bioreactor for the culture of mesenchymal stem cells. In this study, the efficacy of 8 potential coating reagents has been compared in terms of the doubling times of their cell populations, cell morphology, characterization via flow cytometry, and capacity for trilineage differentiation. Human fibronectin (FN), pooled human cryoprecipitate (CPPT), and recombinant human vitronectin (VN) were successful as coating reagents, and each product has advantages in different cell culture contexts. Mesenchymal stem cells harvested from Quantum cultured with each of these 3 compounds as coating reagents all met International Society for Cellular Therapy standards for plastic adherence, surface marker expression, and successful trilineage differentiation. No significant differences were observed among the doubling times from Quantum harvests using FN, CPPT, or VN as coating reagents (P = 0.31). Coating with gelatin, human serum albumin, collagen I, poly‑l‑lysine, and poly‑d‑lysine resulted in significantly lower harvest yield; these agents are not recommended for use as coating reagents in the Quantum system.

Keywords: Automated cell culture; Cell therapy; Extracellular matrix; Hollow-fiber bioreactor; Mesenchymal stromal cell; Regenerative medicine.