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流式分析人脂肪组织中血管周围干细胞含量的方法探究
  • ISSN号:1673-713X
  • 期刊名称:《中国医药生物技术》
  • 时间:0
  • 分类:R733.7[医药卫生—肿瘤;医药卫生—临床医学]
  • 作者机构:[1]解放军总医院骨科研究所,北京100853, [2]兰州军区乌鲁木齐总医院骨科,830000
  • 相关基金:国家自然科学基金(81472092);国家高技术研究发展计划(863计划)(2012AA020502)
中文摘要:

目的建立人脂肪组织中分离血管周围干细胞(PSCs)的方法,并研究其在脂肪组织细胞中所占的比例,为血管周围干细胞作为骨和软骨组织工程新的种子细胞奠定基础。方法取人的脂肪组织分别用 I 型胶原酶和 II 型胶原酶消化得到血管基质成分(SVF),用细胞计数仪及流式细胞仪检测 SVF 中细胞密度、活细胞比例和 PSCs 细胞所占的比例。结果用细胞计数仪分析得出用 II 型胶原酶消化脂肪组织所得到的 SVF 中活细胞比例更高,且差异具有统计学意义(P〈0.01),而细胞密度有增高趋势(P=0.18)。用流式细胞仪检测得出使用 II 型胶原酶消化脂肪组织所得 SVF中 PSCs 比例更高,且差异具有统计学意义(P〈0.01)。使用 I 型胶原酶消化每10 ml 脂肪组织中可以分离得到PSCs 活细胞为(7.98±2.38)×105个,使用 II 型胶原酶消化则每10 ml 脂肪组织中可以分离得到 PSCs 活细胞为(16.68±4.56)×105个。结论使用 II 型胶原酶消化脂肪组织可以得到更多的血管周围干细胞 PSCs,其在脂肪组织中的含量可以满足骨和软骨损伤后自体细胞移植修复的需要。

英文摘要:

Objective To establish the method of sorting out perivascular stem cells (PSCs) from human adipose tissue and study the proportion of these cells in adipose tissue cells. This research is to explore new seed cells for the bone and cartilage tissue engineering. Methods Stromal vascular fraction (SVF) was got from human adipose tissue that was digested by collagenase type I or collagenase type II. The cell density, proportion of living cells and proportion of PSCs in SVF were tested by the cell count and flow cytometry (FCM). Results The proportion of living cells in SVF digested by collagenase type II was much higher through analyzing by the cell count and the difference was statistically significant (P〈0.01). The cellular density in SVF digested by collagenase type II tend to increase (P=0.18). The proportion of PSCs in SVF digested by collagenase type II was much higher through analyzing by the flow cytometry, and the difference was statistically significant (P〈0.01). (7.98 ± 2.38) × 105 PSCs were got in 10 ml human adipose tissue digested by collagenase type I, while (16.68 ± 4.56) × 105 PSCs were got in 10 ml human adipose tissue digested by collagenase type II. Conclusion A higher amount of PSCs can be got from human adipose digested by collagenase type II, and the content of PSCs in the adipose tissue can satisfy the needs of autologous cell transplantation for the bone and cartilage repair.

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期刊信息
  • 《中国医药生物技术》
  • 中国科技核心期刊
  • 主管单位:中华人民共和国卫生部
  • 主办单位:中国医药生物技术协会
  • 主编:蒋建东
  • 地址:北京市天坛西里1号
  • 邮编:100050
  • 邮箱:cmbj01@126.com
  • 电话:010-62115986
  • 国际标准刊号:ISSN:1673-713X
  • 国内统一刊号:ISSN:11-5512/R
  • 邮发代号:80-622
  • 获奖情况:
  • 国内外数据库收录:
  • 被引量:2104