Department of Surgery, Division of Vascular and Endovascular Surgery, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
J Surg Res. 2010 Oct;163(2):e105-12. doi: 10.1016/j.jss.2010.04.025. Epub 2010 May 10.
Most research evaluating adipose-derived stem cells (ASC) uses tissue obtained from young, healthy patients undergoing plastic surgical procedures. Given the propensity of other adult stem cell lines to diminish with increasing patient age and co-morbidities, we assess the availability of ASC in elderly patients undergoing vascular surgical procedures, and evaluate their acquisition of endothelial cell (EC) traits to define their potential use in vascular tissue engineering.
Adipose tissue obtained by liposuction from patients undergoing vascular procedures (n = 50) was digested with collagenase and centrifuged to remove mature adipocytes. The resultant number of cells, defined as the stromal-vascular (SV) pellet, was quantified. Following a 7-d culture period and negative selection for CD31 and CD45, the resultant number of ASC was quantified. After culture in differentiating media (EMG-2), ASCs were tested for the acquisition of endothelial-specific traits (expression of CD31, realignment in shear, cord formation on Matrigel).
The SV pellet contained 2.87 ± 0.34 × 10(5) cells/g fat, and the resultant number of ASCs obtained was 1.41 ± 0.18 × 10(5) cells/g fat. Flow cytometry revealed a homogeneous ASC population (>98% positive for CD13, 29, 90). Advanced age or co-morbidity (obesity, diabetes, renal or peripheral vascular disease) did not significantly alter yield of ASC. After culture in differentiating media (EMG-2), ASCs acquired each of the endothelial-specific traits.
ASC isolation appears independent of age and co-morbidities, and ASCs harvested from patients with vascular disease retain their ability to differentiate into endothelial-like cells. Adipose tissue, therefore, is a practical source of autologous, adult stem cells for vascular tissue engineering.
大多数评估脂肪来源干细胞（ASC）的研究都使用从接受整形手术的年轻、健康患者中获得的组织。鉴于其他成体干细胞系随着患者年龄和合并症的增加而减少的倾向，我们评估了接受血管手术的老年患者中 ASC 的可用性，并评估了它们获得内皮细胞（EC）特征的能力，以确定它们在血管组织工程中的潜在用途。
通过血管手术患者（n=50）的抽脂获得脂肪组织，用胶原酶消化并离心以去除成熟脂肪细胞。剩余的细胞数量定义为基质血管（SV）沉淀，并进行定量。经过 7 天的培养期和 CD31 和 CD45 的阴性选择后，定量 ASC 的数量。在分化培养基（EMG-2）中培养后，测试 ASC 是否获得内皮细胞特异性特征（CD31 表达、在切变下重新排列、在 Matrigel 上形成索状结构）。
SV 沉淀中含有 2.87±0.34×10(5)个细胞/g 脂肪，获得的 ASC 数量为 1.41±0.18×10(5)个细胞/g 脂肪。流式细胞术显示 ASC 群体均匀（>98% 为 CD13、29、90 阳性）。高龄或合并症（肥胖、糖尿病、肾或外周血管疾病）并未显著改变 ASC 的产量。在分化培养基（EMG-2）中培养后，ASC 获得了每种内皮细胞特异性特征。
ASC 的分离似乎与年龄和合并症无关，并且从患有血管疾病的患者中提取的 ASC 保留了分化为内皮样细胞的能力。因此，脂肪组织是用于血管组织工程的自体成体干细胞的实用来源。
