Single-platform quality control assay to quantify multipotential stromal cells in bone marrow aspirates prior to bulk manufacture or direct therapeutic use

Abstract

Absolute quantification of CD45(-/low) CD271(bright) cells was found to be a reliable method of predicting CFU-F yields in BM aspirates. This rapid (< 40 min) procedure could be suitable for intra-operative quality control of BM aspirates prior to volume reduction/direct injection in orthopedics. In the production of culture-expanded MSC, this assay could be used to exclude samples containing low numbers of MSC, resulting in improved consistency and quality of manufactured MSC batches.

Soliciting Strategies for Developing Cell-Based Reference Materials to Advance Mesenchymal Stromal Cell Research and Clinical Translation

“…BMMSCs, bone marrow MSCs; UCB-MSCs, umbilical cord blood MSCs; UCT-MSCs, umbilical cord tissue MSCs; AT-MSCs, adipose tissue MSCs; DP-MSCs, dental pulp MSCs; PMSCs, placental MSCs. Anti-STRO-1, anti-CD146, anti-CD271, anti-nestin positive, CD45-negative cells [22,[24][25][26][27] In vitro trilineage differentiation [18] In vitro immunoplasticity assay of MSCs activated by IFN-g -TNF-a [21] IDO activation in primed MSCs [21,26,[28][29][30] In vitro clonal CFU-F formation efficiency [31] In vivo ossicle formation [22] IFN-g, interferon-gamma; TNF-a, tumor necrosis factor-alpha; IDO, indoleamine 2,3,-dioxygenase; iNOS, inducible nitric oxide synthase; CFU-F, colony-forming unit-fibroblast.…”

Concise Review: Optimizing Expansion of Bone Marrow Mesenchymal Stem/Stromal Cells for Clinical Applications

“…A bone marrow aspirate contains roughly 5-30 3 10 6 MNCs per milliliter and 500-3,000 MSCs per milliliter, the latter determined through CFU-F [11]. Because CFU-F frequency decreases with increasing aspirate volume [95], a 5-ml volume is generally drawn, which may contain only 2,500-6,000 MSCs in total. Thus, the demand for a fast and reliable ex vivo expansion method is necessary.…”

Hierarchy of mesenchymal stem cells: Comparison of multipotentmesenchymal stromal cells with fibroblast colony forming units

“…In order to clarify the differences between stromal precursor cells in CFU-F poor and rich bone marrow samples, they were divided in 2 groups with more and less than median CFU-F concentration. The cumulative production of MMSC from bone marrow with low concentration of CFU-F was significantly lower than in the group with high CFU-F concentration (Figure 4(a)), that align with [14,34]. The expression levels of SPP1 and PPARG were significantly higher and BMP4 was significantly lower in MMSC of CFU-F poor group (Figure 4(b)).…”

“…Sometimes the MMSC content of bone marrow samples is estimated by the classic (CFU-F) assay [13]. MMSC number in the bone marrow is correlated positively with CFU-F concentration [14].…”