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用于脐血和动员外周血单个核细胞来源造血祖细胞体外扩增的全血清剥夺培养基的比较

Comparison of whole serum-deprived media for ex vivo expansion of hematopoietic progenitor cells from cord blood and mobilized peripheral blood mononuclear cells.

作者信息

Sandstrom C E, Collins P C, McAdams T A, Bender J G, Papoutsakis E T, Miller W M

机构信息

Department of Chemical Engineering, Northwestern University, Evanston, IL 60208-3120, USA.

出版信息

J Hematother. 1996 Oct;5(5):461-73. doi: 10.1089/scd.1.1996.5.461.

Abstract

A whole serum-deprived (WSD) medium was developed and optimized for expansion of colony-forming cells (CFC) in cord blood (CB) mononuclear cell (MNC) cultures. This medium was compared with four commercially available WSD media (commercial media), three WSD media whose compositions have been publicly disclosed (public media), two serum-containing media, and two basal media, for cell and CFC expansion in 10-day CB and mobilized peripheral blood (PB) MNC cultures supplemented with interleukin-3 (IL-3), IL-6, and stem cell factor (SCF). Selected WSD media and both serum-containing media gave significant CFC expansion in CBMNC and PBMNC cultures. The serum-containing human long-term medium gave the greatest cell (3.0-fold) and CFC (25-fold) expansions in CBMNC cultures, whereas our medium maintained the most cells (93% of input) and gave the greatest CFC expansion (6.1-fold) for PBMNC cultures. Of the commercial media, Progenitor-34 gave the greatest cell expansion (1.2-fold) and X VIVO-10 gave the greatest CFC expansion (11-fold) for CBMNC cultures, and Progenitor-34 maintained the most cells (83% of input) and gave the greatest CFC expansion (3.1-fold) for PBMNC cultures. Of the public media (including ours), our medium gave the greatest cell (1.4-fold) and CFC (6.1-fold) expansion for CBMNC cultures. Although there were slight correlations between cell and CFC expansion in 10-day CBMNC and PBMNC cultures (r2 of 0.848 and 0.594, respectively), the correlations did not give reliable predictions for medium selection. In addition, the different media favored expansion of different CFC types and performed differently for cultures using different cell sources (CB versus PB). Taken together, these results suggest that media must be carefully screened for the cell source to be cultured and the cell type(s) to be produced (e.g. total cells, CFC).

摘要

开发并优化了一种全血清剥夺(WSD)培养基,用于在脐血(CB)单个核细胞(MNC)培养物中扩增集落形成细胞(CFC)。将该培养基与四种市售WSD培养基(市售培养基)、三种成分已公开的WSD培养基(公开培养基)、两种含血清培养基和两种基础培养基进行比较,用于在添加白细胞介素-3(IL-3)、IL-6和干细胞因子(SCF)的10天CB和动员外周血(PB)MNC培养物中进行细胞和CFC扩增。选定的WSD培养基和两种含血清培养基在CBMNC和PBMNC培养物中均使CFC显著扩增。含血清的人类长期培养基在CBMNC培养物中使细胞扩增最多(3.0倍)和CFC扩增最多(25倍),而我们的培养基在PBMNC培养物中保留了最多的细胞(输入细胞的93%)并使CFC扩增最多(6.1倍)。在市售培养基中,Progenitor-34在CBMNC培养物中使细胞扩增最多(1.2倍),X VIVO-10在CBMNC培养物中使CFC扩增最多(11倍),而Progenitor-34在PBMNC培养物中保留了最多的细胞(输入细胞的83%)并使CFC扩增最多(3.1倍)。在公开培养基(包括我们的培养基)中,我们的培养基在CBMNC培养物中使细胞(1.4倍)和CFC(6.1倍)扩增最多。尽管在10天的CBMNC和PBMNC培养物中细胞和CFC扩增之间存在轻微相关性(r2分别为0.848和0.594),但这些相关性对于培养基选择并不能给出可靠的预测。此外,不同的培养基有利于不同类型CFC的扩增,并且对于使用不同细胞来源(CB与PB)的培养表现不同。综上所述,这些结果表明,对于要培养的细胞来源和要产生的细胞类型(例如总细胞、CFC),必须仔细筛选培养基。

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