AutoMACS Pro Separator from Miltenyi Biotec

January 28, 2013

V.A.Almazov Federal Center for Heart, Blood & Endocrinology, Saint-Petersburg
Research Unit of Cell and Gene Engineering
Department Head

Overall

Performance

Ease-of-Optimization

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MACS-sorted human bone marrow CD34+ cells utilized in cell migration experiment. Migration of CD34+ cells (control sample) from top to the bottom chamber of Transwell system, toward mesenchymal stem cells of healthy and diseased donors.

Company:

Miltenyi Biotec

Product Name:

AutoMACS Pro Separator

Catalog Number:

130-092-545

Info

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This review is for the autoMACS Pro, which has been discontinued and replaced by the autoMACS Neo.

ImageOur team is involved in a number of projects related to adult stem cell biology. Namely, we evaluate key features of stem cells and progenitor cells isolated from bone marrow, adipose tissue, peripheral blood of human and rat, and also from human umbilical cord blood. Key studies include characterization of cell antigen profile (via FACS and ICH), proliferation, induced differentiation, migration, etc. We compare cells isolated from healthy people and those from patients with cardiological (chronic heart failure), endocrinological (diabetes type 2; obesity) and oncohematological (AML, CML, ALL, CLL, MM, MDS, etc.) disorders, in particular. While we possessed cliniMACS unit for clinical applications, and a simple miniMACS set for 1-time experiments, it seemed that we needed more throughput and flexibility in order to fulfill our everyday needs. A robotized system of the preparatory (not clinical) scale seemed to fit perfectly. Briefly, MACS (magnetic activated cell sorting) employs antigen-specific magnetic microbeads to enrich or deplete a heterogeneous cell population of a particular cell type/types. When purchased, it was clearly superior to FACS systems of the time in cell sorting (in cell viability most of all). When loaded with samples, consumables and after being programmed, autoMACS Pro Separator can work in a fully automated manner, decreasing manual labor manyfold. The system represents an impressive combination of mechanics, electronics and fluidics, working in 3D-environment. 

Experimental Design and Results Summary

Application

Most often, we isolate CD34+, CD133+ and CD45+ cells, also perform lineage cell depletion, CD138+ cell isolation, etc. in order to conduct studies in cell proliferation, differentiation, migration and more.

Starting Material

We have experience with autoMACS Pro Separator effectively and flexibly handling a variety of the samples, including peripheral blood, umbilical cord blood and bone marrow.

Protocol Overview

Programs of isolation (single or double load, positive or negative selection) are selected from the wide choice available, based on the sample type, abundance of the cells, etc. For many experiments, we use both positive and negative fractions of the sorting product. Actual samples are prepared following the protocols of the individual kits: generally, there are a few sequential steps of adding beads and reagents, incubating, centrifuging, etc. It may take up to 1.5 hrs to process a set of samples in parallel for the magnetic sorting. Including washing steps, it takes about 10 min per sample to perform the isolation (may be less or longer, depending on the program selected). Finished samples can already be taken off the rack while the sorter keeps working on the next sample in the sequence.

Tips

When using the sorter for large-scale experiments (for many isolations in a row), make sure you have enough reagent in all bottles, for the complete set of isolations programmed. If the sorter will run out of the buffer (for example) in the middle of the run, it will ruin the sample. Since programming is so simple, we recommend setting up shorter sets (no more than 3 isolations in a row), and checking on the volume of reagents in between. Despite much tubing inside the sorter, we have never observed a leak, but make sure you use the wrench to check and tighten all connections weekly. Chill racks hold the temperature for about 40 min: if you have many experiments, you can order additional chill rack or two, of the size you use most (we use 15 ml for almost all of our isolations).

Results Summary

The sorter splits the sample into negative and positive fractions of the defined volume, with good purity and fine cell viability. Quite a small number of cells loaded for the sorting are lost: i.e., are not uploaded into ‘+’ and ‘–‘ fractions.

Features Summary

The sorter can work even with very rare cells, which is indeed useful. Sample overload is generally not an issue, since the upper margin of the working range (max. number of labeled cells and of total cells) is rather high for autoMACS columns. In a number of the protocols (see the particular kits), double/sequential selection for 2 markers is possible. After the cell count, cell sorting product quality is checked via FACS analysis, or cells are utilized directly: for cell-cell interaction, cell migration studies, or any other downstream applications.

Additional Notes

Handling of the instrument is quite easy: it does not require complex procedures often, column change sequence (performed biweekly) is simple, and the rest comes to supplying fresh bottles of reagents and emptying the waste tank. Programs and actions are selected and launched via the touchpad, interface is well-designed.

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MACS-sorted human bone marrow CD34+ cells utilized in cell migration experiment. Migration of CD34+ cells (control sample) from top to the bottom chamber of Transwell system, toward mesenchymal stem cells of healthy and diseased donors.

Summary

The Good

Automation increases throughput and reduces variability. A choice from pre-loaded programs is just great.

The Bad

The system is very sensitive to the temperature; it breaks if air conditioning fails. The manual is far from perfect.

The Bottom Line

A marvelous piece of mechanics, electronics and fluidics. Great balance of throughput and ease-of-use. Perfect for experiments of a preparatory scale.

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