Osteoporosis is a disease characterized by low bone mineral density and overall weakening of bone. It shows sex association, affecting post-menopausal women at higher rates than men. It is also is age specific, showing increased prevalence with aging in both sexes. Due to the increased mortality and severe disruptions in quality of life caused by osteoporosis-related fractures, the disease poses a serious public health concern (Sözen, Özışık, & Başaran, 2017). While osteoporosis is known to be related to environmental factors and associated lifestyle factors, recent genome-wide association studies (GWAS) have found dozens of loci associated with low bone mineral density and increased fracture risk, necessitating further study into specific causal genes (Sabik & Farber, 2017; Zheng, Spector, & Richards, 2011). Bone self-renewal, throughout life, is facilitated by resident mesenchymal stem cell (MSC) populations found within bone tissues, which are capable of osteogenic differentiation. Establishment of reliable cell culture methods for the growth and differentiation of bone marrow MSCs is a pre-requisite to carrying out comparative population-level studies of MSCs focused on assessing the interactions between underlying genetic and co-morbidity features that affect their osteogenic potential. Culture media is often supplemented with fetal bovine serum (FBS), containing essential components that promote cell growth. However, use of FBS has led to issues in controllability and reproducibility of data from cultures supplemented with FBS because of its poorly-defined nature and variations in composition from sources of FBS. In this study, the efficacy of bone marrow-derived MSC (BMSC) cultures grown in a commercially available artificial media was compared to cells grown in media containing FBS, and their growth and osteogenic differentiation capacity were compared.
Human MSCs were obtained from the acetabular reaming samples from 10 patients undergoing total hip arthroplasty at Boston Medical Center. We processed the raw materials to remove tissue and adipose materials, and seeded using the loose cellular component of the bone marrow in media supplemented with FBS and commercially available serum-free media, Mesencult-ACF (Stem Cell Technologies, Vancouver, Canada). Adherent cell MSC populations were selected over a seven-day growth period. Osteogenic differentiation was initiated using supplementation with Dexamethasone, ascorbate, and -glycerol phosphate. Cultured cells were grown in both media conditions for 21 days after osteogenesis was initiated. We examined cellular DNA content for cell growth, alkaline phosphatase (ALP) for osteogenic differentiation and calcium content for mineralization and terminal osteocyte differentiation.
We did not find significant differences between the groups for average DNA, ALP or calcium content (p=0.167, p=0.139, p=0.291). We did find significant differences within each subject between groups for nearly every subject and across all measurements. We did find significant differences in the DNA content and ALP content of the FBS supplemented media group between subjects (p=0.045, p=0.020). Findings suggest Mesencult-ACF is a suitable alternative to media supplemented with FBS for promoting growth and differentiation of BMSCs.
| Identifer | oai:union.ndltd.org:bu.edu/oai:open.bu.edu:2144/36522 |
| Date | 12 June 2019 |
| Creators | Diaz, Stephanie |
| Contributors | Gerstenfeld, Louis C., Divieti Pajevic, Paola |
| Source Sets | Boston University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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