Researchers from the Critical Analytics for Manufacturing Personalized-Medicine (CAMP) Interdisciplinary Research Group (IRG) of Singapore-MIT Alliance for Research and Technology (SMART), MIT’s analysis enterprise in Singapore, in collaboration with the Agency for Science, Technology and Research (A*STAR) Bioprocessing Technology Institute (BTI) and National University Health System (NUHS), have developed a groundbreaking expertise able to extracting mesenchymal stem cells (MSCs) instantly from pure bone marrow – often known as bone marrow aspirate (BMA), a pivotal supply of MSCs – with out dilution.

Using a world-first steady sorting approach on a multi-chip Deterministic Lateral Displacement (DLD) microfluidic platform, the brand new technique doubles the amount of MSCs obtained from bone marrow samples and shortens the time taken to round 20 minutes. It additionally reduces donor discomfort as a consequence of much less bone marrow being extracted, hurries up cell manufacturing, and simplifies cell remedy manufacturing. This development represents a big step towards extra accessible and environment friendly superior medical remedies that use MSCs – together with remedy for osteoarthritis, autoimmune and infectious illnesses, and neurological problems.

Cell remedy is a discipline of medication the place cells are used as residing medicine to combat illnesses or restore and change broken cells. Advances in regenerative drugs and immunotherapy have benefited numerous sufferers; they provide quite a few new remedy options to beforehand intractable illnesses, with tons of extra within the developmental pipeline that provides new hope to sufferers. However, acquiring high-quality uncooked supplies – on this case, MSCs – has lengthy been a hurdle in cell remedy manufacturing, as conventional isolation strategies comparable to centrifugation are inefficient and complicated. In addition to an extended processing time of round two to 3 hours, present strategies end in low yield as a consequence of challenges comparable to osmotic stress and complicated workflows. State-of-the-art sorting strategies comparable to fluorescence-activated cell sorting (FACS) depend on pricey antibodies and complicated preparation, posing main limitations for manufacturing of those residing medicines.

In a paper titled “Scalable mesenchymal stem cell enrichment from bone marrow aspirate using DLD microfluidic sorting”, lately printed within the journal Lab on a Chip, SMART researchers have pioneered a revolutionary stem cell sorting platform, addressing the challenges of large-scale cell sorting and manufacturing. Using DLD microfluidic expertise, a label-free cell sorting technique which distinguishes between stem cells and blood cells, the platform processed small bone marrow samples (2.5 mL) in simply 20 minutes with double the stem cell yield in comparison with conventional strategies, and bypasses pricey reagents and complicated processes.

In this technique pioneered by SMART, human bone marrow samples that arrive on the laboratory endure a easy filtration step to take away undesirable cells and tissues that would impede the chip. Samples are then loaded onto SMART’s sorting platform, and cells of curiosity (MSCs) are mechanically sorted and picked up in outlet reservoirs. These collected cells are then mixed right into a vial for additional processing and quantification as wanted.

This progressive breakthrough in cell sorting makes use of microfluidic applied sciences, leveraging cells’ pure properties and eliminating the need for labelling. With standard strategies, cells are sorted utilizing fluorescent or magnetic tags to label sure mobile options. This is difficult as these labels may intrude with subsequent evaluation and testing, or worse injury the cells. In comparability, passive strategies just like the DLD technique developed at SMART are user-friendly, mild on cells and simply built-in into medical pattern processing workflows. MSCs are very delicate to its exterior surroundings, and perturbations may alter the biology in unanticipated methods. 

“This novel platform provides a fresh perspective for stem cell sorting through a more efficient, label-free and, importantly, seamless process integration into current industrial manufacturing pipeline. Our research team at SMART is excited about the possibilities this technology brings to the field of stem cell research and therapy. The successful demonstration of this technology gives us greater confidence to venture into other bioprocessing applications such as leukopheresis with great potential for clinical impact. This will significantly accelerate the development of cutting-edge treatments and improve accessibility for cell therapy,” stated Mr Nicholas Tan, Research Engineer at SMART CAMP and lead creator of the paper.

“Even though DLD cell sorting has previously been demonstrated, what is new in this work is that we were able to deploy the technique at a sufficiently high processing flow rate to impact real-world stem cell manufacturing workflow. Biomanufacturing and bioprocessing are areas in which I see much potential for applying microfluidics technology to improve overall efficiency and reduce the cost significantly,” stated Professor Jongyoon Han, Co-Lead Principal Investigator at SMART CAMP, Professor of Biological Engineering and Electrical Engineering at MIT and corresponding creator of the paper.

Future efforts will deal with refining the expertise by evaluating the standard of sorted MSCs from human bone marrow samples utilizing strategies comparable to reverse transcription polymerase chain response (RT-PCR) and differentiation assays. Simultaneously, CAMP is working in direction of rising sorting pace and determination, whereas refining the system’s design for portability and user-friendliness, and rising throughput to 10 ml per minute.

Our progressive strategy marks a paradigm shift in cell sorting, a key technique of cell remedy. By harnessing microfluidic applied sciences to capitalise on cells’ intrinsic properties, now we have eradicated the necessity for cumbersome and costly labelling strategies. It not solely streamlines the sorting course of but in addition ensures extra correct and dependable leads to medical analysis. This breakthrough, pushed by our dedication to advancing scientific frontiers, heralds a milestone within the realm of mobile research.”

Dr Kerwin Kwek, Research Scientist, SMART CAMP

Dr Kerwin Kwek can also be co-lead creator of the paper.

The analysis is carried out by SMART and supported by the National Research Foundation (NRF) Singapore beneath its Campus for Research Excellence And Technological Enterprise (CREATE) programme. National University Hospital offered the bone marrow pattern, and A*STAR BTI carried out the traditional MSC isolation technique which CAMP’s expertise was benchmarked towards.