Label-free mesenchymal stem cell enrichment from bone marrow samples by inertial microfluidics†,Analytical Methods

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Label-free mesenchymal stem cell enrichment from bone marrow samples by inertial microfluidics†
Analytical Methods ( IF 2.7 ) Pub Date : 2018-01-09 00:00:00 , DOI: 10.1039/c7ay02500a
Lap Man Lee _{1,

2,

3} , Jenna M. Rosano _{1,

2,

3} , Yi Wang _{3,

4,

5,

6} , George J. Klarmann _{3,

7,

8,

9} , Charles J. Garson _{1,

2,

3} , Balabhaskar Prabhakarpandian _{1,

2,

3} , Kapil Pant _{1,

2,

3} , Luis M. Alvarez _{3,

7,

8,

9,

10} , Eva Lai _{3,

11,

12,

13,

14}

Affiliation

Isolation of pure populations of mesenchymal stem cells from bone marrow aspirate is a critical need in regenerative medicine such as orthopedic and cartilage reconstruction with important clinical and therapeutic implications. Currently available stem cell isolation systems mainly rely on intrusive immuno-labeling techniques. Mesenchymal stem cells in bone marrow samples are typically larger than other cells, which can be used as a distinctive biophysical cue for non-invasive isolation. In this work, a spiral-shaped inertial microfluidic sorter was developed to isolate mesenchymal stem cells from mouse bone marrow samples with minimal sample preparation steps. To characterize the sorting performance, cultured mesenchymal stem cells were spiked into tissue-digested mouse bone marrow cells. At a flow rate of 1.6 mL min⁻¹, an average enrichment of 6.0× and a recovery of 73% were demonstrated. About 3 × 10⁶ tissue-digested bone marrow cells can be processed in 1 minute with a single microfluidic run. The recovered mesenchymal stem cells after microfluidic sorting retained high (>95%) viability and similar immuno-phenotype expressions and multi-potencies in tri-differentiation lineages to unprocessed cells. This rapid, label-free and non-invasive inertial microfluidic sorter has practical applications in target stem cell enrichment in stem cell therapy.

中文翻译：

惯性微流控从骨髓样品中富集无标记的间充质干细胞†

从骨髓穿刺物中分离出纯净的间充质干细胞群体是再生医学（如骨科和软骨重建）的关键需求，具有重要的临床和治疗意义。当前可用的干细胞分离系统主要依靠侵入性免疫标记技术。骨髓样品中的间充质干细胞通常比其他细胞大，可以用作非侵入性分离的独特生物物理线索。在这项工作中，开发了一种螺旋形惯性微流体分选仪，以最少的样品制备步骤从小鼠骨髓样品中分离出间充质干细胞。为了表征分选性能，将培养的间充质干细胞掺入组织消化的小鼠骨髓细胞中。在1.6 mL min ^-1的流速下，平均富集度为6.0倍，回收率达73％。一次微流操作可在1分钟内处理大约3×10 ^6个组织消化的骨髓细胞。微流分选后恢复的间充质干细胞保留了高（> 95％）的生存力，并且在三分化谱系中具有与未处理细胞相似的免疫表型表达和多潜能。这种快速，无标签且无创的惯性微流分类器在干细胞治疗中的目标干细胞富集中具有实际应用。

更新日期：2018-01-09

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