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研究经细胞因子和基质细胞衍生因子 1 混合物处理的扩增人 CD34 细胞的干细胞归巢和自我更新标记基因特征。

Study of stem cell homing & self-renewal marker gene profile of expanded human CD34 cells manipulated with a mixture of cytokines & stromal cell-derived factor 1.

机构信息

Chiplunkar Laboratory, Advanced Centre for Treatment, Research & Education in Cancer, Tata Memorial Centre, Navi Mumbai; Homi Bhabha National Institute (HBNI), Dr. LH Hiranandani Hospital, Mumbai, India.

Bone Marrow Transplant Unit, Department of Medical Oncology, Advanced Centre for Treatment, Research & Education in Cancer, Tata Memorial Centre, Navi Mumbai, India.

出版信息

Indian J Med Res. 2017 Jul;146(1):56-70. doi: 10.4103/ijmr.IJMR_1319_15.

DOI:10.4103/ijmr.IJMR_1319_15
PMID:29168461
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5719609/
Abstract

BACKGROUND & OBJECTIVES: Next generation transplantation medicine aims to develop stimulating cocktail for increased ex vivo expansion of primitive hematopoietic stem and progenitor cells (HSPC). The present study was done to evaluate the cocktail GF (Thrombopoietin + Stem Cell factor + Flt3-ligand) and homing-defining molecule Stromal cell-derived factor 1 (SDF1) for HSPC ex vivo expansion.

METHODS

Peripheral blood stem cell (n=74) harvests were analysed for CD34hiCD45lo HSPC. Immunomagnetically enriched HSPC were cultured for eight days and assessed for increase in HSPC, colony forming potential in vitro and in vivo engrafting potential by analyzing human CD45+ cells. Expression profile of genes for homing and stemness were studied using microarray analysis. Expression of adhesion/homing markers were validated by flow cytometry/ confocal microscopy.

RESULTS

CD34hiCD45lo HSPC expansion cultures with GF+SDF1 demonstrated increased nucleated cells (n=28, P+ cells (n=8, P=0.021) and increased colony forming units (cfu) compared to unstimulated and GF-stimulated HSPC. NOD-SCID mice transplanted with GF+SDF1-HSPC exhibited successful homing/engraftment (n=24, PInterpretation & conclusions: Cocktail of cytokines and SDF1 showed good potential to successfully expand HSPC which exhibited enhanced ability to generate multilineage cells in short-term and long-term repopulation assay. This cocktail-mediated stem cell expansion has potential to obviate the need for longer and large volume apheresis procedure making it convenient for donors.

摘要

背景与目的

下一代移植医学旨在开发刺激鸡尾酒，以增加原始造血干细胞和祖细胞（HSPC）的体外扩增。本研究旨在评估 HSPC 体外扩增的细胞因子鸡尾酒 GF（血小板生成素+干细胞因子+Flt3 配体）和归巢定义分子基质细胞衍生因子 1（SDF1）。

方法

分析外周血干细胞（n=74）采集物中的 CD34hiCD45lo HSPC。用免疫磁珠富集 HSPC，培养 8 天，并通过分析人 CD45+细胞来评估 HSPC 增加、体外和体内集落形成潜能以及植入潜能。使用微阵列分析研究归巢和干性基因的表达谱。通过流式细胞术/共聚焦显微镜验证粘附/归巢标记物的表达。

结果

GF+SDF1 的 CD34hiCD45lo HSPC 扩增培养显示有核细胞（n=28，P<0.001）、CD34+细胞（n=8，P=0.021）和 CFU 增加，与未刺激和 GF 刺激的 HSPC 相比。用 GF+SDF1-HSPC 移植的 NOD-SCID 小鼠表现出成功的归巢/植入（n=24，P<0.001）。

解释和结论

细胞因子和 SDF1 的鸡尾酒显示出良好的潜力，可以成功地扩增 HSPC，这些 HSPC 表现出增强的在短期和长期重建造血实验中生成多谱系细胞的能力。这种鸡尾酒介导的干细胞扩增有可能避免对更长和更大体积的单采程序的需求，使供体更加方便。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0d7/5719609/865c72f35468/IJMR-146-56-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0d7/5719609/39141d6c0e8a/IJMR-146-56-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0d7/5719609/731742e50135/IJMR-146-56-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0d7/5719609/4cce8f04cfba/IJMR-146-56-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0d7/5719609/0c930352ad85/IJMR-146-56-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0d7/5719609/12ba87767c02/IJMR-146-56-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0d7/5719609/865c72f35468/IJMR-146-56-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0d7/5719609/39141d6c0e8a/IJMR-146-56-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0d7/5719609/731742e50135/IJMR-146-56-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0d7/5719609/4cce8f04cfba/IJMR-146-56-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0d7/5719609/0c930352ad85/IJMR-146-56-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0d7/5719609/12ba87767c02/IJMR-146-56-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0d7/5719609/865c72f35468/IJMR-146-56-g009.jpg

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