Human Adipose-Derived Mesenchymal Stromal Cells Exhibit High HLA-DR Levels and Altered Cellular Characteristics under a Xeno-free and Serum-free Condition,Stem Cell Reviews and Reports

当前位置： X-MOL 学术 › Stem Cell Rev and Rep. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Human Adipose-Derived Mesenchymal Stromal Cells Exhibit High HLA-DR Levels and Altered Cellular Characteristics under a Xeno-free and Serum-free Condition
Stem Cell Reviews and Reports ( IF 4.5 ) Pub Date : 2021-09-11 , DOI: 10.1007/s12015-021-10242-7
Phuong T M Dam ₁ , Van T Hoang ₂ , Hue Thi Hong Bui ₁ , Le Minh Hang ₁ , Duc M Hoang ₂ , Hoang Phuong Nguyen ₂ , Ha Thi Lien ₁ , Huong Thi Thanh Tran ₁ , Xuan-Hung Nguyen ₁ , Liem Nguyen Thanh ₂

Affiliation

Background

We have observed an increased expression of negative markers in some clinical-grade, xeno- and serum-free cultured adipose-derived mesenchymal stem/stromal cell (ADMSC) samples. It gave rise to concern that xeno- and serum-free conditions might have unexpected effects on human ADMSCs. This study aims to test this hypothesis for two xeno- and serum-free media, PowerStem MSC1 media (PS) and StemMACS MSC Expansion Media (SM), that support the in vitro expansion of ADMSCs.

Methods

We investigated the expression of negative markers in 42 clinical-grade ADMSC samples expanded in PS. Next, we cultured ADMSCs from seven donors in PS and SM and examined their growth and colony-forming ability, surface marker expression, differentiation, cell cycle and senescence, as well as genetic stability of two passages representing an early and late passage for therapeutic MSCs.

Results

15 of 42 clinical-grade PS-expanded ADMSC samples showed an increased expression of negative markers ranging from 2.73% to 34.24%, which positively correlated with the age of donors. This rise of negative markers was related to an upregulation of Human Leukocyte Antigen – DR (HLA-DR). In addition, the PS-cultured cells presented decreased growth ability, lower frequencies of cells in S/G2/M phases, and increased ß-galactosidase activity in passage 7 suggesting their senescent feature compared to those grown in SM. Although MSCs of both PS and SM cultures were capable of multilineage differentiation, the PS-cultured cells demonstrated chromosomal abnormalities in passage 7 compared to the normal karyotype of their SM counterparts.

Conclusions

These findings suggest that the SM media is more suitable for the expansion of therapeutic ADMSCs than PS. The study also hints a change of ADMSC features at more advanced passages and with increased donor’s age. Thus, it emphasizes the necessity to cover these aspects in the quality control of therapeutic MSC products.

Graphical abstract

中文翻译：

人类脂肪来源的间充质基质细胞在无异种物质和无血清条件下表现出高 HLA-DR 水平和改变的细胞特征

背景

我们观察到一些临床级、异种和无血清培养的脂肪来源的间充质干/基质细胞 (ADMSC) 样本中阴性标志物的表达增加。这引起了人们对异种和无血清条件可能对人类 ADMSC 产生意想不到的影响的担忧。本研究旨在为支持 ADMSCs 体外扩增的两种异种和无血清培养基 PowerStem MSC1 培养基 (PS) 和 StemMACS MSC 扩增培养基 (SM) 检验这一假设。

方法

我们研究了在 PS 中扩展的 42 个临床级 ADMSC 样本中阴性标志物的表达。接下来，我们在 PS 和 SM 中培养了来自 7 个供体的 ADMSCs，并检查了它们的生长和集落形成能力、表面标志物表达、分化、细胞周期和衰老，以及代表治疗性 MSCs 早期和晚期传代的两个传代的遗传稳定性.

结果

42 个临床级 PS 扩增的 ADMSC 样本中有 15 个显示阴性标志物的表达增加，范围从 2.73% 到 34.24%，这与供体的年龄呈正相关。这种阴性标志物的上升与人类白细胞抗原 - DR (HLA-DR) 的上调有关。此外，与在 SM 中生长的细胞相比，PS 培养的细胞生长能力下降，S/G2/M 期细胞频率降低，并且在第 7 代中 ß-半乳糖苷酶活性增加，表明它们具有衰老特征。尽管 PS 和 SM 培养的 MSC 都能够进行多向分化，但与 SM 对应物的正常核型相比，PS 培养的细胞在第 7 代中表现出染色体异常。