Preliminary evaluation of a highly automated instrument for the selection of CD34+ cells from mobilized peripheral blood stem cell concentrates

Abstract

Background Cell selection is an important part of manufacturing cellular therapies. A new highly automated instrument, the CliniMACS Prodigy, was evaluated for the selection of CD34+ cells from mobilized peripheral blood stem cell (PBSC) concentrates using monoclonal antibodies conjugated to para-magnetic particles. Methods PBSCs were collected by apheresis from 36 healthy subjects given G-CSF or G-CSF plus plerixafor. CD34+ cells from 11 PBSC concentrates were isolated with the automated CliniMACS Prodigy and 25 with the semi-automated CliniMACS Plus Instrument. Results The proportion of CD34+ cells in the selected products obtained with the two instruments was similar; 93.6±2.6% for the automated and 95.7±3.3% for the semi-automated instrument (p > 0.05). The recovery of CD34+ cells from PBSC concentrates was less for the automated than the semi-automated instrument (51.4±8.2% versus 65.1±15.7%; p=0.019). The selected products from both instruments contained few and similar quantities of platelets and red blood cells. The depletion of CD3+ cells was less with the automated instrument (4.34±0.2 log depletion versus 5.20±0.35 log depletion; p < 1 × 10−6). Removal of platelets from PBSC concentrates by washing was associated with better CD34+ cell recovery. We explored the reasons for lower CD34+ cell recovery by the Prodigy and found that the non-selected cells for the Prodigy contained more platelets than those for the CliniMACS Plus. Conclusions CD34+ cells can be effectively selected from mobilized PBSC concentrates with the CliniMAC Prodigy, but the recovery of CD34+ cells and depletion of CD3+ cells was lower than with the semi-automated CliniMACS Plus Instrument.

Genetic Engineering and Manufacturing of Hematopoietic Stem Cells

“…It has been reported that a medium recovery of 71% and medium purity of 97% of CD34 + cells, with a median of 0.04% residual CD3 + cells could be achieved from either BM or MPB by using the CliniMACS device 23 . Similar results were obtained with the automated CliniMACS Prodigy, although CD34 + cell recovery and depletion of CD3 + cells may be lower than with the semi-automated CliniMACS Plus instrument 24, 25…”

Prospective Evaluation of a Practical Guideline for Managing Positive Sterility Test Results in Cell Therapy Products

“…As pioneers in gene therapy manufacturing since the 1990s 22 , we introduced closed-system processing (from flask to bag-based cell culture) as a means to reduce product contamination 23 . These methods continue to be used and optimized for recent immunotherapies, including the manufacture of clinical grade CAR T-cells, TCR- transduced T-cells, mesenchymal stromal cells, and mutivirus specific T-cells 24–26 .…”

Transplantation of gene-modified haematopoietic stem cells: Application and clinical considerations

“…These devices are certified for cell therapy applications and globally accepted by regulatory authorities. [14,15] However, the system is costly and has low throughput, which will have its challenges if we are to foresee roll-out of ex vivo gene-modified cell therapies for diseases such as HIV and sickle cell disease where the patient burden is in the millions and is often in lower-to-middle income countries. There is thus an opportunity to address these limitations with more efficient process developments, lower cost and higher throughput equipment solutions.…”