Mass Production of Early-Stage Bone-Marrow-Derived Mesenchymal Stem Cells of Rat Using Gelatin-Coated Matrix

Abstract

Although preparation of early-stage bone-marrow-derived mesenchymal stem cells (BM-MSCs) is critical for successful cell transplantation therapy, no culture system offers a sufficient number of early-stage BM-MSCs for cell transplantation. Accordingly, we developed a culture system capable of producing a large number of early-stage BM-MSCs by using gelatin-coated matrix. The greatest retrieval and proliferation rates of the earliest-stage rat BM-MSCs were detected in bone-marrow-derived cells cultured on 1% (wt/v) gelatin-coated matrix, which showed significantly greater colony forming unit-fibroblast number, diameter, and total cell number. Moreover, continuous culture of the earliest-stage BM-MSCs on 1% (wt/v) gelatin-coated matrix resulted in a maximum of 21.2 ± 2.7 fold increase in the cumulative total number of early-stage BM-MSCs at passage 5. BM-MSCs generated in large quantities due to a reduced doubling time and an increased yield of cell population in S/G2/M phase showed typical fibroblast-like morphology and no significant differences in BM-MSC-related surface marker expression and differentiation potential, except for an increased ratio of differentiation into a neurogenic lineage. The use of gelatin-coated matrix in the retrieval and culture of BM-MSCs contributes greatly to the effective isolation and mass production of early-stage BM-MSCs.

GFP Labeling and Hepatic Differentiation Potential of Human Placenta-Derived Mesenchymal Stem Cells

“…To calculate the population doubling time (PDT), 3 × 10 3 cells/cm 2 were seeded in 24-well plates (Nunclon™ Delta Surface). The doubling time of the GFP + hPMSCs and control cells were calculated according to the formula: PDT=t log2/(logN t − logN 0 ), where t is the time to confluence, N t is the number of cells at the end of the growth period, and N 0 is the initial number of cells [10].…”

Boron containing poly-(lactide-co-glycolide) (PLGA) scaffolds for bone tissue engineering

“…Cells were washed with PBS and analyzed using Becton Dickinson FACS Calibur flow cytometry system (Becton Dickinson, San Jose, CA, USA). In addition, cells (P3) were differentiated into adipogenic [29], osteogenic [30] and chondrogenic [31] lineages to confirm mesenchymal stem cell characteristics according to the protocols described previously with minor modifications. Differentiation medium compositions are given in Supplementary Table 1.…”

Small RNA sequencing reveals a novel tsRNA‐06018 playing an important role during adipogenic differentiation of hMSCs

“…Importantly, peak tsRNA expression was evident on day 7 of adipogenesis, leading us to focus on this time point, which has also been previously shown to be a critical period that regulates the fate of differentiating pre-adipocytic cells. 36,37 We therefore selected the differentially expressed tsRNAs on days 7 and 14 to analyse their expression trends during adipogenic differentiation (Figure 2A,B), revealing trends consistent with the proportion and reads of these tsRNAs. Interestingly, on day 7 of F I G U R E 6 tsRNA-06018 targets the 3′-UTR of STC2.…”