CD146富集在牙髓息肉来源干细胞纯化中的作用

Role of CD146 Enrichment in Purification of Stem Cells Derived from Dental Pulp Polyp.

作者信息

Tavangar Maryam Sadat, Hosseini Seyed-Mojtaba, Dehghani-Nazhvani Ali, Monabati Ahmad

机构信息

Department of Operative Dentistry, Dental School, Shiraz University of Medical Sciences, Shiraz, Iran.

Student Research Committee,Cellular and Molecular Research Club, Shiraz University of Medical Sciences, Shiraz, Iran; ; Cardiovascular Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.

出版信息

Iran Endod J. 2017 Winter;12(1):92-97. doi: 10.22037/iej.2017.19.

Abstract

INTRODUCTION

Hyperplastic pulpitis (pulp polyp) tissues contains cells with stem cell properties similar to that of the dental pulp stem cells (DPSCs). It has also been shown that CD146 enrichment can homogenize the cultures of DPSCs and enhance the colony forming potentials of their cultures. This study determines whether CD146 enrichment can help purifying the stem cells from heterogeneous cultures of the pulp polyp derived stem cells (PPSCs).

METHODS AND MATERIALS

Healthy dental pulps and pulp polyp tissues were enzymatically digested and the harvested single cells were sorted according to the presence of CD146 marker. The sorted cells were seeded directly for colony forming unit (CFU) assays of the negative and positive portions. Flowcytometric antigen panel and differentiation assays were used to see if these cells conform with mesenchymal stems cells (MSCs) definition. Differences between the between groups was assessed using independent t-test. The level of significance was set at 0.05.

RESULTS

Normal pulp tissue derived cells formed higher colonies (42.5±16.8 per 10 cells) than the pulp polyp (17.75±8.9 per 10 cells) (=0.015). The CD146 positive portion of the polyp derived cells formed an average of 91.5±29.7 per 10 cells per CFU. On the other hand, CD146 negative portion did not show any colonies (<0.001). Both resources showed cells with flowcytometric antigen panel and differentiation potentials conforming to MSC definition.

CONCLUSION

The entire CFU of PPSCs were formed within CD146 enriched portion. It seems that CD146 enrichment may reduce the number of possible fibroblasts of the pulp polyps and may further homogenize the culture of the PPSCs.

摘要

引言

增生性牙髓炎(牙髓息肉)组织中含有具有与牙髓干细胞(DPSCs)相似干细胞特性的细胞。研究还表明,富集CD146可使DPSCs培养物同质化,并增强其培养物的集落形成能力。本研究旨在确定富集CD146是否有助于从牙髓息肉衍生干细胞(PPSCs)的异质培养物中纯化干细胞。

方法与材料

对健康牙髓和牙髓息肉组织进行酶消化,收获的单细胞根据CD146标记物的存在进行分选。将分选后的细胞直接接种,用于阴性和阳性部分的集落形成单位(CFU)测定。采用流式细胞术抗原分析和分化分析来观察这些细胞是否符合间充质干细胞(MSCs)的定义。使用独立t检验评估组间差异。显著性水平设定为0.05。

结果

正常牙髓组织来源的细胞形成的集落(每10个细胞42.5±16.8个)比牙髓息肉(每10个细胞17.75±8.9个)多(P=0.015)。息肉来源细胞的CD146阳性部分平均每10个细胞每CFU形成91.5±29.7个集落。另一方面,CD146阴性部分未显示任何集落(P<0.001)。两种来源的细胞均显示出具有符合MSCs定义的流式细胞术抗原分析和分化潜能。

结论

PPSCs的整个CFU在CD146富集部分形成。似乎富集CD146可能减少牙髓息肉中可能的成纤维细胞数量,并可能进一步使PPSCs培养物同质化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/087c/5282388/149b052de351/iej-12-092-g001.jpg

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