一种新型人颗粒细胞分离纯化方案的建立。
Development of a novel protocol for isolation and purification of human granulosa cells.
机构信息
Department of Obstetrics & Gynecology, Division of Reproductive Endocrinology and Infertility, University of Texas Medical Branch, Galveston, TX, USA.
出版信息
J Assist Reprod Genet. 2012 Jun;29(6):547-56. doi: 10.1007/s10815-012-9739-5. Epub 2012 Mar 10.
Abstract
PURPOSE
To develop an optimal method of isolation and purification of human granulosa cells from ovarian follicular fluid.
METHODS
Follicular fluid was collected from patients undergoing oocyte retrieval. A series of isolation and purification techniques was performed, involving density gradient centrifugation and use of different antibody-bead complexes.
RESULTS
The highest percent yield of live purified granulosa cells came from density gradient centrifugation using sucrose polymer followed by positive selection of granulosa cells using primary antibody to MISRII and secondary antibody coupled to iron oxide beads.
CONCLUSIONS
A novel protocol for granulosa cell purification has been developed yielding samples that are largely free of nondesirable cells. This protocol provides a purification solution, especially for patient samples that have significant RBC contamination.
摘要
目的
开发一种从卵巢卵泡液中分离和纯化人颗粒细胞的最佳方法。
方法
从接受卵母细胞采集的患者中收集卵泡液。进行了一系列的分离和纯化技术，包括密度梯度离心和使用不同的抗体-珠复合物。
结果
使用蔗糖聚合物进行密度梯度离心，然后使用针对 MISRII 的初级抗体和与氧化铁珠偶联的二级抗体对颗粒细胞进行阳性选择，得到的活纯化颗粒细胞的最高百分比产量。
结论
已经开发出一种新的颗粒细胞纯化方案，得到的样品几乎不含不需要的细胞。该方案提供了一种纯化解决方案，特别是对于红细胞污染严重的患者样本。
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2
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Reprod Biol Endocrinol. 2010 Apr 2;8:32. doi: 10.1186/1477-7827-8-32.
3
Differential gene expression in human granulosa cells from recombinant FSH versus human menopausal gonadotropin ovarian stimulation protocols.
Reprod Biol Endocrinol. 2010 Mar 12;8:25. doi: 10.1186/1477-7827-8-25.
4
The role of autophagy in follicular development and atresia in rat granulosa cells.
Fertil Steril. 2010 May 15;93(8):2532-7. doi: 10.1016/j.fertnstert.2009.11.021. Epub 2010 Feb 10.
5
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Environ Health Perspect. 2010 Mar;118(3):400-6. doi: 10.1289/ehp.0901161. Epub 2009 Oct 22.
6
Flow cytometric assay for the simultaneous determination of residual white blood cells, red blood cells, and platelets in fresh-frozen plasma: validation and two years' experience.
Transfusion. 2009 Jun;49(6):1195-204. doi: 10.1111/j.1537-2995.2008.02079.x. Epub 2009 Feb 6.
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