使用Histopaque从兔血中分离单核细胞。

Use of Histopaque for isolating mononuclear cells from rabbit blood.

作者信息

Feldman D L, Mogelesky T C

机构信息

Department of Pharmacology and Chemotherapy, Hoffmann-La Roche Inc., Nutley, NJ 07110.

出版信息

J Immunol Methods. 1987 Sep 24;102(2):243-9. doi: 10.1016/0022-1759(87)90083-4.

DOI:10.1016/0022-1759(87)90083-4

PMID:3655375

Abstract

This paper describes the use of Histopaque (Hp) density gradient medium and its advantage for separating mononuclear cells (mnc) from rabbit blood. Hp solutions of density (d) = 1.083 g/ml, 1.119 g/ml and 1:1 mixture of these solutions (final d = 1.103 g/ml) were compared to Ficoll-Paque (Fp, d = 1.077 g/ml) and Lymphopaque (d = 1.086 g/ml). The average leukocyte recovery was: Fp = 26% and Lp = 17%, while that with Hp was: Hp d = 1.083: 41%; Hp d = 1.119: 43%; Hp d = 1.103: 57%. Optimum mnc recovery (76%) was obtained with Hp d = 1.103. Average mnc purity for the various media was: Fp = 88% mnc; Lp = 91%mnc; Hp d = 1.083: 94% mnc; Hp d = 1.119: 93%; Hp d = 1.103: 94% mnc. Contamination was mainly from basophils. Viability was greater than 95% in all cases. Thus, Hp density gradient media provide increased recoveries of mnc from rabbit blood compared to Fp and Lp, while purity is not affected. Rabbit mnc appear to be denser than human mnc, which are recovered in greater numbers using Fp.

摘要

本文描述了Histopaque（Hp）密度梯度介质的使用及其从兔血中分离单核细胞（mnc）的优势。将密度（d）=1.083 g/ml、1.119 g/ml的Hp溶液以及这些溶液的1:1混合物（最终d = 1.103 g/ml）与Ficoll-Paque（Fp，d = 1.077 g/ml）和Lymphopaque（d = 1.086 g/ml）进行比较。白细胞平均回收率为：Fp = 26%，Lp = 17%，而使用Hp的回收率为：Hp d = 1.083：41%；Hp d = 1.119：43%；Hp d = 1.103：57%。使用Hp d = 1.103可获得最佳单核细胞回收率（76%）。各种介质的单核细胞平均纯度为：Fp = 88%单核细胞；Lp = 91%单核细胞；Hp d = 1.083：94%单核细胞；Hp d = 1.119：93%；Hp d = 1.103：94%单核细胞。污染主要来自嗜碱性粒细胞。所有情况下活力均大于95%。因此，与Fp和Lp相比，Hp密度梯度介质可提高从兔血中回收单核细胞的效率，同时不影响纯度。兔单核细胞似乎比人单核细胞密度更大，人单核细胞使用Fp回收数量更多。