Balakumar Balasubramanian, Rajagopal Karthikeyan, Madhuri Vrisha
Paediatric Orthopaedics Unit, Department of Orthopaedics, Christian Medical College, Vellore, India.
Centre for Stem Cell Research, Christian Medical College, Vellore, India.
Indian J Med Res. 2016 Dec;144(6):831-837. doi: 10.4103/ijmr.IJMR_8_15.
BACKGROUND & OBJECTIVES: Human bone marrow is rich in various growth factors which may support the chondrocyte growth. This study was conducted to compare the culture characteristics of human growth plate chondrocyte in foetal bovine serum (FBS) and human autologous bone marrow extract (BME) in monolayer culture.
Iliac crest apophyseal cartilage was harvested from four donors, aged between two and nine years, undergoing hip surgery. Chondrocytes were propagated under two culture conditions, with 10 per cent FBS and 10 per cent autologous BME harvested from the same donors. Cells were harvested at 7, 14 and 21 days to assess viability, morphology, cell count and immunocytochemistry.
With an initial seeding density of 2500 cells/cm 2 , the average yield in monolayer cultured with FBS was 3.35 × 10 5 , 5.9 × 10 5 , 14.1 × 10 5 and BME was 0.66 × 10 5 , 1.57 × 10 5 and 3.48 × 10 5 at 7, 14 and 21 days, respectively. Viability was 98.21 per cent with FBS and 97.45 per cent with BME at 21 days. In BME supplemented cultures, hyaline phenotype was maintained up to 21 days. The yield was higher in the FBS supplemented group; however, the phenotype could not be maintained by the FBS group as long as BME group.
INTERPRETATION & CONCLUSIONS: Autologous BME was found to be a safer alternative to FBS for human studies. BME could maintain the hyaline phenotype for a longer time. Ways to enhance the cell yield needs to be explored in future studies.
人骨髓富含多种可支持软骨细胞生长的生长因子。本研究旨在比较人胎儿牛血清(FBS)和人自体骨髓提取物(BME)在单层培养中对人生长板软骨细胞的培养特性。
从4名年龄在2至9岁接受髋关节手术的供体获取髂嵴骨骺软骨。软骨细胞在两种培养条件下增殖,分别添加10%来自相同供体的FBS和10%自体BME。在第7、14和21天收获细胞,评估细胞活力、形态、细胞计数和免疫细胞化学。
初始接种密度为2500个细胞/cm²时,在FBS单层培养中,第7、14和21天的平均产量分别为3.35×10⁵、5.9×10⁵、14.1×10⁵,而在BME培养中分别为0.66×10⁵、1.57×10⁵和3.48×10⁵。第21天时,FBS组细胞活力为98.21%,BME组为97.45%。在添加BME的培养物中,透明软骨表型维持至21天。FBS添加组产量更高;然而,FBS组不能像BME组那样长时间维持表型。
自体BME被发现是人体研究中FBS的更安全替代品。BME可更长时间维持透明软骨表型。未来研究需要探索提高细胞产量的方法。