Jurgens Wouter J F M, Oedayrajsingh-Varma Maikel J, Helder Marco N, Zandiehdoulabi Behrouz, Schouten Tabitha E, Kuik Dirk J, Ritt Marco J P F, van Milligen Florine J
Department of Plastic, Reconstructive, and Hand Surgery, VU University Medical Center, Amsterdam, The Netherlands.
Cell Tissue Res. 2008 Jun;332(3):415-26. doi: 10.1007/s00441-007-0555-7. Epub 2008 Apr 1.
The stromal vascular fraction (SVF) of adipose tissue contains an abundant population of multipotent adipose-tissue-derived stem cells (ASCs) that possess the capacity to differentiate into cells of the mesodermal lineage in vitro. For cell-based therapies, an advantageous approach would be to harvest these SVF cells and give them back to the patient within a single surgical procedure, thereby avoiding lengthy and costly in vitro culturing steps. However, this requires SVF-isolates to contain sufficient ASCs capable of differentiating into the desired cell lineage. We have investigated whether the yield and function of ASCs are affected by the anatomical sites most frequently used for harvesting adipose tissue: the abdomen and hip/thigh region. The frequency of ASCs in the SVF of adipose tissue from the abdomen and hip/thigh region was determined in limiting dilution and colony-forming unit (CFU) assays. The capacity of these ASCs to differentiate into the chondrogenic and osteogenic pathways was investigated by quantitative real-time polymerase chain reaction and (immuno)histochemistry. A significant difference (P = 0.0009) was seen in ASC frequency but not in the absolute number of nucleated cells between adipose tissue harvested from the abdomen (5.1 +/- 1.1%, mean +/- SEM) and hip/thigh region (1.2 +/- 0.7%). However, within the CFUs derived from both tissues, the frequency of CFUs having osteogenic differentiation potential was the same. When cultured, homogeneous cell populations were obtained with similar growth kinetics and phenotype. No differences were detected in differentiation capacity between ASCs from both tissue-harvesting sites. We conclude that the yield of ASCs, but not the total amount of nucleated cells per volume or the ASC proliferation and differentiation capacities, are dependent on the tissue-harvesting site. The abdomen seems to be preferable to the hip/thigh region for harvesting adipose tissue, in particular when considering SVF cells for stem-cell-based therapies in one-step surgical procedures for skeletal tissue engineering.
脂肪组织的基质血管成分(SVF)含有大量多能脂肪组织来源的干细胞(ASC),这些干细胞在体外具有分化为中胚层谱系细胞的能力。对于基于细胞的治疗方法,一种有利的途径是收集这些SVF细胞,并在单一外科手术过程中将其回输给患者,从而避免冗长且昂贵的体外培养步骤。然而,这要求SVF分离物含有足够数量能够分化为所需细胞谱系的ASC。我们研究了ASC的产量和功能是否受最常用于采集脂肪组织的解剖部位(腹部和髋部/大腿区域)的影响。通过极限稀释和集落形成单位(CFU)测定法确定了来自腹部和髋部/大腿区域的脂肪组织的SVF中ASC的频率。通过定量实时聚合酶链反应和(免疫)组织化学研究了这些ASC分化为软骨生成和成骨途径的能力。从腹部采集的脂肪组织(5.1±1.1%,平均值±标准误)和髋部/大腿区域采集的脂肪组织(1.2±0.7%)之间,ASC频率存在显著差异(P = 0.0009),但有核细胞的绝对数量没有差异。然而,在源自这两个组织的CFU中,具有成骨分化潜能的CFU频率是相同的。培养时,获得了具有相似生长动力学和表型的均匀细胞群体。两个组织采集部位的ASC之间在分化能力上未检测到差异。我们得出结论,ASC的产量取决于组织采集部位,但每体积有核细胞的总量以及ASC的增殖和分化能力并不取决于组织采集部位。在考虑用于骨骼组织工程的一步式外科手术中基于干细胞治疗的SVF细胞时,腹部似乎比髋部/大腿区域更适合采集脂肪组织。