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脐血单个核细胞体外诱导分化为粒系细胞的研究

OBJECTIVE: To explore an optimal method for granulocyte cell production from umbilical cord blood mononuclear cells. METHODS: Erythrocytes were precipitated by hydroxyethyl starch. Mononuclear cells were isolated through Ficoll density gradient centrifugation. Different media, additives and cultivat...

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Formato: Online Artículo Texto
Lenguaje:English
Publicado: Editorial office of Chinese Journal of Hematology 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7342961/
https://www.ncbi.nlm.nih.gov/pubmed/28655099
http://dx.doi.org/10.3760/cma.j.issn.0253-2727.2017.06.013
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collection PubMed
description OBJECTIVE: To explore an optimal method for granulocyte cell production from umbilical cord blood mononuclear cells. METHODS: Erythrocytes were precipitated by hydroxyethyl starch. Mononuclear cells were isolated through Ficoll density gradient centrifugation. Different media, additives and cultivation model were chosen for granulocyte induction. Cell morphology was observed by microscopy, and cell phenotype was detected by flow cytometry. The CD18 expression of granulocytes was tested by immunofluorescence assay, and phagocytosis test was executed as well. RESULTS: Compared to fetal bovine serum (FBS) treatment group, cell viability, counts and differentiation rate of granulocytes induced by X-VIVO™ 15 combined with TPO, SCF, G-CSF but without FBS were superior. And X-VIVO™ 15 medium was better than SCGM medium at effectiveness and cost. Using two-stage mode of hematopoietic stem cell expansion followed by granulocyte induction with X-VIVO™ 15 combining TPO, SCF and G-CSF, cell proliferation was nearly 132 times at day 21. Flow cytometry showed that the differentiation was lagged in 2-stage mode than in direct induction mode, CD15 expression was (69.60± 1.06) % vs (97.73±0.39) %; Wright-Giemsa staining demonstrated mature granulocytes; immunofluorescence showed the expression of lysosomal proteins CD18. A strong phagocytic function of mature granulocytes was demonstrated by phagotrophic efficiency of (51.43±0.05) %. And granulocyte had chemotaxis ability under the role of chemotactic factor IL-8. CONCLUSION: Optimized culture media and cultivation mode are achieved for functional granulocytes induction in vitro.
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spelling pubmed-73429612020-07-16 脐血单个核细胞体外诱导分化为粒系细胞的研究 Zhonghua Xue Ye Xue Za Zhi 论著 OBJECTIVE: To explore an optimal method for granulocyte cell production from umbilical cord blood mononuclear cells. METHODS: Erythrocytes were precipitated by hydroxyethyl starch. Mononuclear cells were isolated through Ficoll density gradient centrifugation. Different media, additives and cultivation model were chosen for granulocyte induction. Cell morphology was observed by microscopy, and cell phenotype was detected by flow cytometry. The CD18 expression of granulocytes was tested by immunofluorescence assay, and phagocytosis test was executed as well. RESULTS: Compared to fetal bovine serum (FBS) treatment group, cell viability, counts and differentiation rate of granulocytes induced by X-VIVO™ 15 combined with TPO, SCF, G-CSF but without FBS were superior. And X-VIVO™ 15 medium was better than SCGM medium at effectiveness and cost. Using two-stage mode of hematopoietic stem cell expansion followed by granulocyte induction with X-VIVO™ 15 combining TPO, SCF and G-CSF, cell proliferation was nearly 132 times at day 21. Flow cytometry showed that the differentiation was lagged in 2-stage mode than in direct induction mode, CD15 expression was (69.60± 1.06) % vs (97.73±0.39) %; Wright-Giemsa staining demonstrated mature granulocytes; immunofluorescence showed the expression of lysosomal proteins CD18. A strong phagocytic function of mature granulocytes was demonstrated by phagotrophic efficiency of (51.43±0.05) %. And granulocyte had chemotaxis ability under the role of chemotactic factor IL-8. CONCLUSION: Optimized culture media and cultivation mode are achieved for functional granulocytes induction in vitro. Editorial office of Chinese Journal of Hematology 2017-06 /pmc/articles/PMC7342961/ /pubmed/28655099 http://dx.doi.org/10.3760/cma.j.issn.0253-2727.2017.06.013 Text en 2017年版权归中华医学会所有 http://creativecommons.org/licenses/by-nc-sa/3.0/ This work is licensed under a Creative Commons Attribution 3.0 License (CC-BY-NC). The Copyright own by Publisher. Without authorization, shall not reprint, except this publication article, shall not use this publication format design. Unless otherwise stated, all articles published in this journal do not represent the views of the Chinese Medical Association or the editorial board of this journal.
spellingShingle 论著
脐血单个核细胞体外诱导分化为粒系细胞的研究
title 脐血单个核细胞体外诱导分化为粒系细胞的研究
title_full 脐血单个核细胞体外诱导分化为粒系细胞的研究
title_fullStr 脐血单个核细胞体外诱导分化为粒系细胞的研究
title_full_unstemmed 脐血单个核细胞体外诱导分化为粒系细胞的研究
title_short 脐血单个核细胞体外诱导分化为粒系细胞的研究
title_sort 脐血单个核细胞体外诱导分化为粒系细胞的研究
topic 论著
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7342961/
https://www.ncbi.nlm.nih.gov/pubmed/28655099
http://dx.doi.org/10.3760/cma.j.issn.0253-2727.2017.06.013
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