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不同代次牙髓干细胞分泌细胞因子的比较研究及其向神经细胞分化的潜能。

A comparative study on the secretion of various cytokines by pulp stem cells at different passages and their neurogenic potential.

机构信息

Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Hamburg 20246, Germany.

Department of Oral and Maxillofacial Surgery, Guiyang Hospital of Stomatology, Guiyang, 050017, PR China.

出版信息

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2022 May;166(2):161-167. doi: 10.5507/bp.2021.058. Epub 2021 Nov 5.

Abstract

AIMS

By measuring the extent of cytokines secreted by human dental pulp stem cells (hDPSCs) from passages 2 through 10, the optimal passage of hDPSCs was determined. This offers a potential theoretical basis for the treatment of neurological disorders.

METHOD

After isolation and culture of hDPSCs from human teeth, the morphological features of the cells were observed under an inverted microscope. hDPSCs were identified by their immunophenotypes and their multiple differentiation capability. Cytokine concentrations secreted in the supernatants at passages 2-10 were detected by ELISA.

RESULTS

hDPSCs were viewed as fusiform or polygonal in shape, with a bulging cell body, homogenized cytoplasm, and a clear nucleus. Moreover, they could differentiate into neuroblasts in vitro. hDPSCs at passage 3 were positive for CD29 (91.5%), CD73 (94.8%) and CD90 (96.7%), but negative for the hematopoietic markers CD34 (0.13%). ELISA results showed that hDPSCs at passage 3 had the highest secretion levels of vascular endothelial growth factor (VEGF), brain-derived neurotrophic factor (BDNF), and nerve growth factor (NGF), with the highest secretion level of Neurotrophin-3 (NT-3) being at passage 2.

CONCLUSION

hDPSCs have steady biological features of stem cells and exhibit optimal proliferation potential. hDPSCs at different passages have different capacities in the secretion of VEGF, BDNF, NGF, and NT-3. In conclusion cytokines secreted by hDPSCs may prove to be appropriate in the treatment of neurological diseases.

摘要

目的

通过测量人牙髓干细胞(hDPSCs)第 2 至 10 代分泌的细胞因子的程度,确定 hDPSCs 的最佳传代数。这为治疗神经退行性疾病提供了潜在的理论基础。

方法

从人牙中分离和培养 hDPSCs 后,在倒置显微镜下观察细胞的形态特征。通过免疫表型和多向分化能力鉴定 hDPSCs。通过 ELISA 检测上清液中第 2-10 代分泌的细胞因子浓度。

结果

hDPSCs 呈梭形或多角形,细胞体饱满,细胞质均匀,核清晰。此外,它们可以在体外分化为神经母细胞。第 3 代 hDPSCs 表达 CD29(91.5%)、CD73(94.8%)和 CD90(96.7%),但不表达造血标志物 CD34(0.13%)。ELISA 结果表明,第 3 代 hDPSCs 分泌血管内皮生长因子(VEGF)、脑源性神经营养因子(BDNF)和神经生长因子(NGF)的水平最高,第 2 代 hDPSCs 分泌神经营养因子-3(NT-3)的水平最高。

结论

hDPSCs 具有稳定的干细胞生物学特征,表现出最佳的增殖潜力。不同传代的 hDPSCs 在 VEGF、BDNF、NGF 和 NT-3 的分泌能力上存在差异。总之,hDPSCs 分泌的细胞因子可能适用于治疗神经退行性疾病。

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