Radhi Noor, Paul Ayamita, Muelbert Mariana, Toldi Gergely
Department of Perinatal Science, Liggins Institute, University of Auckland, Auckland, New Zealand.
J Hum Lact. 2025 May;41(2):263-271. doi: 10.1177/08903344251316491. Epub 2025 Feb 18.
There is increasing interest in better understanding the immune cell composition of human milk and how these cells interact with neonatal immune development. However, consistent methods for immune cell isolation from human milk are lacking.
Our aim was to systematically compare available cell isolation techniques to isolate T lymphocytes from human milk samples.
This repeated measures study design compared three cell isolation methods using human milk samples: (1) centrifugation, (2) immunomagnetic bead isolation, and (3) a combination of both methods. We assessed the proportion and viability of CD3+, CD4+, CD25+ and regulatory T cells using flow cytometry in isolated cells to compare the performance of these isolation methods.
Immunomagnetic separation is a feasible method to isolate T lymphocytes in human milk, similar to blood. It improves target cell enrichment and cell viability compared to centrifugation, which may be an advantage when the goal is to characterize rare cell types or when cells are further used in functional assays. No excess cell activation (CD25 positivity) was observed with the use of magnetic beads.
Immunomagnetic separation of human milk T lymphocytes may have advantages over centrifugation depending on the intended downstream use of cells.
人们越来越关注更好地了解人乳中的免疫细胞组成以及这些细胞如何与新生儿免疫发育相互作用。然而，目前缺乏从人乳中分离免疫细胞的一致方法。
我们的目的是系统比较从人乳样本中分离T淋巴细胞的现有细胞分离技术。
本重复测量研究设计比较了三种使用人乳样本的细胞分离方法：（1）离心法，（2）免疫磁珠分离法，以及（3）两种方法的组合。我们使用流式细胞术评估分离细胞中CD3 +、CD4 +、CD25 +和调节性T细胞的比例和活力，以比较这些分离方法的性能。
免疫磁珠分离是一种从人乳中分离T淋巴细胞的可行方法，类似于从血液中分离。与离心法相比，它提高了目标细胞的富集率和细胞活力，当目标是鉴定稀有细胞类型或细胞进一步用于功能测定时，这可能是一个优势。使用磁珠未观察到过度的细胞活化（CD25阳性）。
根据细胞预期的下游用途，人乳T淋巴细胞的免疫磁珠分离可能比离心法具有优势。
