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Cells were passaged when colonies reached 80% confluency

Cells were passaged when colonies reached 80% confluency. marrow had been more responsive to chondrogenic induction, forming superior cartilaginous tissue in vitro, compared to those isolated from femoral head. (4) The identification of discrete progenitor populations in bone marrow by current cell-surface marker profiling is not predictive for subsequently derived in vitro BMSC cultures. Overall, the iliac crest and the vertebral body offer a Aescin IIA more reliable tissue source of stromal progenitor cells for cartilage repair strategies compared to femoral head. = ?0.17, < 0.34; PDT: = ?0.23, = 0.15)), (C,D) bone marrow source (fh: femoral head, ic: iliac crest, vb: vertebral body) and gender did not affect MSC yield or PDT. KruskalCWallis statistics with Dunns multiple comparison test was performed to test for differences in yield/seeded and PDT, respectively, between different tissue sources. = 33. Table 1 Demographical information from patients where bone marrow samples were acquired. The total yield of mononuclear cells (MNCs) isolated by density centrifugation is shown. Bone marrow-derived stromal cells (BMSC) yield at p0/MNC originally seeded and the population doubling time (p0Cp1) of tissue-culture plastic selected and monolayer expanded stromal cell populations. = 11 (E); = 13 (F,G). 2.3. Progenitor Abundance in the Aescin IIA Freshly Isolated Mononuclear Cell Fraction Is Not Predictive of the Derived Monolayer Expanded BMSC Populace The abundance of progenitor populations within the MNC fraction from different bone marrow sources was assessed (Physique 3ACD). Inter-donor variability was apparent with no statistically significant difference in progenitor abundance, as determined by our marker panels, between different anatomical sites. We next investigated whether MNC fractions made up of a higher frequency of putative progenitor populations would give rise to higher BMSC yields following monolayer growth. The abundance of CD45?CD34?CD146+, CD146+/NG2+, or CD45?CD34?CD73+ populations in the freshly isolated MNC fraction did not correlate with the yield of BMSCs at p0 (Physique 3ECG). No significant correlations were apparent between the frequency of progenitors in the freshly isolated MNC fraction and the population doubling time of the subsequently derived stromal populace in the analyzed samples, which were mainly derived from Rabbit polyclonal to ALS2CL vertebral body (Physique 3ICK). We observed a trend between the abundance of CD146+NG2+ cells Aescin IIA and the population doubling time; however, this was not statistically significant (= 0.09, Figure 3J). We further analyzed CD45 (marker of the haematopoietic lineage) to test whether the abundance of haematopoietic cells in the initial MNC populace might affect the growth kinetics of BMSCs. We found a pattern of a negative correlation of the abundance of CD45+ leukocytes in the MNC fraction with the BMSC yield in p0, which was also not statistically significant (= 0.10, Figure 3H). No correlation was observed between the abundance of CD45+ in the MNC fraction and the population doubling time (Physique 3L). Open in a separate window Physique 3 Abundance of putative progenitor populations from different bone marrow sources and correlation analyses. Comparison of Aescin IIA the frequency of MSC populations, identified as (A) Aescin IIA CD45?CD34?CD146+ (= 12), (B) CD146+NG2+ (= 13), or (C) CD45?CD34?CD73+ (= 17) and (D) CD45+ leukocytes (= 17) in bone marrow (BM)-derived MNCs from different sources. Frequency of all cell populations was variable in individual donors but not significantly different in-between tissue sources (fh = femoral head, ic = iliac crest, vb = vertebral body),.


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