Tissue Engineering Group, Department of Orthopaedic Surgery, Faculty of Medicine, National Orthopaedic Centre of Excellence for Research and Learning, University of Malaya, 50603, Kuala Lumpur, Malaysia.
J Mater Sci Mater Med. 2011 May;22(5):1343-56. doi: 10.1007/s10856-011-4294-7. Epub 2011 Apr 2.
The use of mesenchymal stem cells (MSCs) in tissue repair and regeneration despite their multipotentiality has been limited by their cell source quantity and decelerating proliferative yield efficiency. A study was thus undertaken to determine the feasibility of using microcarrier beads in spinner flask cultures for MSCs expansion and compared to that of conventional monolayer cultures and static microcarrier cultures. Isolation and characterization of bone marrow derived MSCs were conducted from six adult New Zealand white rabbits. Analysis of cell morphology on microcarriers and culture plates at different time points (D0, D3, D10, D14) during cell culture were performed using scanning electron microscopy and bright field microscopy. Cell proliferation rates and cell number were measured over a period of 14 days, respectively followed by post-expansion characterization. MTT proliferation assay demonstrated a 3.20 fold increase in cell proliferation rates in MSCs cultured on microcarriers in spinner flask as compared to monolayer cultures (p < 0.05). Cell counts at day 14 were higher in those seeded on stirred microcarrier cultures (6.24 ± 0.0420 cells/ml) × 10(5) as compared to monolayer cultures (0.22 ± 0.004 cells/ml) × 10(5) and static microcarrier cultures (0.20 ± 0.002 cells/ml) × 10(5). Scanning electron microscopy demonstrated an increase in cell colonization of the cells on the microcarriers in stirred cultures. Bead-expanded MSCs were successfully differentiated into osteogenic and chondrogenic lineages. This system offers an improved and efficient alternative for culturing MSCs with preservation to their phenotype and multipotentiality.
尽管间充质干细胞 (MSCs) 具有多能性，但由于其细胞来源数量有限和增殖效率降低，其在组织修复和再生中的应用受到限制。因此，进行了一项研究，以确定在搅拌瓶培养中使用微载体珠来扩大 MSCs 的可行性，并将其与传统的单层培养和静态微载体培养进行比较。从六只成年新西兰白兔中分离和鉴定骨髓来源的 MSCs。在细胞培养过程中的不同时间点 (D0、D3、D10、D14)，使用扫描电子显微镜和明场显微镜分析微载体和培养板上的细胞形态。在 14 天的时间内分别测量细胞增殖率和细胞数量，然后进行扩展后特征分析。MTT 增殖测定显示，与单层培养相比，在搅拌瓶中的微载体上培养的 MSCs 的细胞增殖率提高了 3.20 倍 (p < 0.05)。在第 14 天，接种于搅拌微载体培养物中的细胞数量较高 (6.24 ± 0.0420 个/ml) × 10(5)，与单层培养物 (0.22 ± 0.004 个/ml) × 10(5) 和静态微载体培养物 (0.20 ± 0.002 个/ml) × 10(5)。扫描电子显微镜显示，搅拌培养物中细胞在微载体上的细胞定植增加。扩展后的 MSCs 成功分化为成骨细胞和软骨细胞谱系。该系统为培养 MSCs 提供了一种改进和有效的替代方法，可保持其表型和多能性。
