Manufacturing Differences Affect Human Bone Marrow Stromal Cell Characteristics and Function: Comparison of Production Methods and Products from Multiple Centers

Abstract

Human bone marrow stromal cells (BMSCs, also known as bone marrow-derived mesenchymal stem cells) are manufactured using many different methods, but little is known about the spectrum of manufacturing methods used and their effects on BMSC characteristics and function. Seven centers using, and one developing, Good Manufacturing Practices (GMP) processes were surveyed as to their production methods. Among the seven centers, all used marrow aspirates as the starting material, but no two centers used the same manufacturing methods. Two to four BMSC lots from each center were compared using global gene expression. Among the twenty-four BMSC lots from the eight centers intra-center transcriptome variability was low and similar among centers. Principal component analysis and unsupervised hierarchical clustering analysis separated all the lots from five centers into five distinct clusters. BMSCs from six of the eight centers were tested for their ability to form bone and support hematopoiesis by in vivo transplantation (defining features of BMSCs). Those from all six centers tested formed bone, but the quantity formed was highly variable and BMSCs from only three centers supported hematopoiesis. These results show that differences in manufacturing resulted in variable BMSC characteristics including their ability to form bone and support hematopoiesis.

Mesenchymal stem cells: Cell therapy and regeneration potential

“…Furthermore, processing of BM after collection could also affect the quality and quantity of isolated MSCs. In a comparative study, BM‐MSCs isolated from different good manufacturing practices centers yielded MSCs with different properties, such as the ability to support the hematopoiesis microenvironment and bone formation in vivo (Liu et al, ). Such discrepancies in behavior can limit the autologous and allogenic use of BM‐MSCs .…”

The mesenchymoangioblast, mesodermal precursor for mesenchymal and endothelial cells

“…This includes significant inconsistency among MSC preparations due to donor to donor variability, differences in tissue source, isolation and cell culture techniques, and potential contaminations of mesenchymal cell preparations with non-mesenchymal cells (macrophages, endothelial cells and others). In addition, limited expansion potential of MSCs from adult tissues and lack of markers to define composition and biological function of MSC preparations hamper production of cellular preparations with reproducible pharmacological effects [74,75,72,76,77]. …”

Bone tissue engineering: Adult stem cells in combination with electrospun nanofibrous scaffolds

“…They are spindle‐shaped nucleated cells with multi‐lineage differentiation potential and immune‐modulating ability which most broadly studied for therapeutic potentials. Indeed, BM‐MSCs have been considered as the gold standard for clinical applications (Batsali et al, ), because they are easily extractable, survive better after transplantation and show weak immunogenicity (Husak & Dworzak, ; Liu et al, ; Shin et al, ; Tsuji et al, ; Zeng et al, ). In fact, BM‐MSCs are represented as the cells of origin of osteosarcoma while they are capable of dividing into one of several mesenchymal phenotypes such as osteoblasts, chondrocytes, myocytes, marrow stromal cells, tendon‐ligament fibroblasts, and adipocytes (Caplan & Dennis, ; Tsuji et al, ; He et al, ).…”