Aspiration to obtain osteoblast progenitor cells from human bone marrow: The influence of aspiration volume
Aspiration to obtain osteoblast progenitor cells from human bone marrow: The influence of aspiration volume
复制标题
DOI：
10.2106/00004623-199711000-00012
复制
发表时间：
1997-11-01
影响因子：
5.3
通讯作者：
Easley, K
中科院分区：
文献类型：
--
作者：
Muschler, GF;Boehm, C;Easley, K
骨髓含有成骨细胞祖细胞，可以通过抽吸获得，并且似乎来自多能结缔组织干细胞群体。当在体外培养条件下，促进成骨细胞表型，成骨细胞祖细胞增殖，形成表达碱性磷酸酶的细胞集落，随后，一个成熟的成骨phenotype.We评估的骨髓样本中有核细胞的数量与抽吸从前髂嵴的32例无全身性疾病。共有19名男性患者和13名女性患者;平均年龄为41岁（范围：14 - 77岁）。还通过将骨髓来源的细胞置于组织培养基中并计数碱性磷酸酶阳性集落形成单位的数量来确定成骨细胞祖细胞的患病率和浓度。为了评估抽吸体积的影响，进行了两个连续实验。在第一个实验中，比较了一毫升和两毫升的抽吸体积。在第二个实验中，比较了2毫升和4毫升的抽吸量，骨髓样本中碱性磷酸酶阳性集落形成单位的平均患病率为每百万个有核细胞36个（95%置信区间为28 ~ 47）;从2毫升抽吸物中平均获得2400个碱性磷酸酶阳性集落形成单位。在这些骨髓样本中，碱性磷酸酶阳性集落形成单位的数量在患者之间存在显著差异（p < 0.001）。70%的这种患病率的变化是由于患者之间的变化，20%是由于之间的变化抽吸。碱性磷酸酶阳性的菌落形成单位的数量随着抽吸量的增加而增加。然而，外周血的污染也随着抽吸量的增加而增加。从1到4毫升的抽吸量的增加造成了大约50%的碱性磷酸酶阳性的菌落形成单位的最终浓度在平均samples.Clinical RELEVANCE：在这些数据的基础上，我们建议，当骨髓获得与抽吸用作骨移植，从任何一个网站的抽吸量不应大于两毫升。由于骨髓样本被外周血稀释，较大的体积会降低成骨祖细胞的浓度。我们估计，四个一毫升吸出物将提供碱性磷酸酶阳性集落形成单位的数量几乎是一个四毫升吸出物的两倍。此外，这些数据证实，人类在骨髓的细胞构成和成骨细胞祖细胞的普遍性方面彼此显著不同。需要进一步的研究来确定骨髓中碱性磷酸酶阳性集落形成单位的数量或流行率是否是自体骨或骨髓移植物疗效的决定因素，并确定碱性磷酸酶阳性集落形成单位的数量和功能如何作为年龄、绝经状态和选定疾病等因素的函数而变化。
Bone marrow contains osteoblast progenitor cells that can be obtained with aspiration and appear to arise from a population of pluripotential connective-tissue stem cells. When cultured in vitro under conditions that promote an osteoblastic phenotype, osteoblast progenitor cells proliferate to form colonies of cells that express alkaline phosphatase and, subsequently, a mature osteoblastic phenotype.We evaluated the number of nucleated cells in bone-marrow samples obtained with aspiration from the anterior iliac crest of thirty-two patients without systemic disease. There were nineteen male patients and thirteen female patients; the mean age was forty-one years (range, fourteen to seventy-seven years). The prevalence and concentration of the osteoblast progenitor cells also were determined, by placing the bone-marrow-derived cells into tissue-culture medium and counting the number of alkaline phosphatase-positive colony-forming units. In order to assess the effect of aspiration volume, two sequential experiments were performed. In the first experiment, aspiration volumes of one and two milliliters were compared. In the second experiment, aspiration volumes of two and four milliliters were compared.The mean prevalence of alkaline phosphatase-positive colony-forming units in the bone-marrow samples was thirty-six per one million nucleated cells (95 per cent confidence interval, 28 to 47); a mean of 2400 alkaline phosphatase-positive colony-forming units was obtained from a two-milliliter aspirate. There was a significant difference among the patients with respect to the number of alkaline phosphatase-positive colony-forming units in these bone-marrow samples (p < 0.001). Seventy per cent of this variation in the prevalence was due to variation among patients, and 20 per cent was due to variation among aspirates.The number of alkaline phosphatase-positive colony-forming units in the aspirate increased as the aspiration volume increased. However, contamination by peripheral blood also increased as the aspiration volume increased. An increase in the aspiration volume from one to four milliliters caused a decrease of approximately 50 per cent in the final concentration of alkaline phosphatase-positive colony-forming units in an average sample.CLINICAL RELEVANCE: On the basis of these data, we recommend that, when bone marrow is obtained with aspiration for use as a bone graft, the volume of aspiration from any one site should not be greater than two milliliters. A larger volume decreases the concentration of osteoblast progenitor cells because of dilution of the bone-marrow sample with peripheral blood. We estimate that four one-milliliter aspirates will provide almost twice the number of alkaline phosphatase-positive colony-forming units as will one four-milliliter aspirate. In addition, these data confirm that humans differ significantly from one another with respect to the cellularity of bone marrow and the prevalence of osteoblast progenitor cells. Additional studies are necessary to determine if the number or prevalence of alkaline phosphatase-positive colony-forming units in bone marrow is a determining factor in the efficacy of an autogenous bone or bone-marrow graft and to ascertain how the number and function of alkaline phosphatase-positive colony-forming units may change as a function of factors such as age, menopausal status, and selected diseases.