ALDHbr cells were sorted from fresh bone marrow without prior stimulation, post-processing or culture. Approximately 180 mL (±10mL) of bone marrow was aspirated from the iliac crests from both cell and placebo group participants. In order to isolate ALDHbr cells for cell group participants, the marrow was shipped via courier service to one of two cell processing facilities. Ten patients had cells processed at Aldagen facilities in Durham, NC) and 27 patients had cells processed at Baylor Center for Cell and Gene Therapy (Houston, TX). The product was prepared by enriching mononuclear cells from the marrow using a Sepax device (Biosafe, Geneva, Switzerland), staining the primitive stem cells for the intracellular enzyme ALDH and aseptically sorting using a FACS Aria cell sorter (Becton Dickenson, San Jose, CA). The product was returned by courier service to the clinical center for administration within 96 hours of marrow aspiration. The placebo product consisted of phenol red-free CellGro SCGM serum-free medium (CellGenix Technologie Transfer GmbH, Frieburg, Germany) supplemented with 1% human serum albumin (Flexbumin, Baxter, IL). Both study products (cells and placebo) were identical in appearance.
Flexbumin
Manufactured by Baxter
Sourced in Israel
Flexbumin is a laboratory equipment product designed for albumin separation and purification. It utilizes a flexible membrane technology to efficiently extract and concentrate albumin from biological samples.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using flexbumin
1
ALDHbr Cell Isolation from Bone Marrow
2
GIP and Xen Peptide Synthesis
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3
Expansion of HBsAg-Specific T Cells
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The patient´s peripheral blood mononuclear cells were collected by leukocyte apheresis. Cells were sorted by magnetic beads carrying CD4 and CD8 antibodies (Miltenyi Biotec) on a CliniMACS® Plus Instrument under good-manufacturing-practice (GMP) conditions. Subsequently, the sorted CD4 + and CD8 + T cells were activated by Transact microspheres presenting CD3 and CD28 antibodies (Miltenyi Biotec) and cultured in Prime-XV T-cell medium (Irvine Scientific) with 400 IU/ml IL-2 (Shandong Quangang Co.).
A GMP-grade lentivirus (produced by WuXi AppTec.) encoding the HBsAg-specific TCR gene was added to activated T cells on day 1. Cells were cultured at 37 °C for 8 to 12 days in a closed bag system (Cytiva) on a Xuri Cell Expansion System W25 (Cytiva). Cells were harvested by 300xg centrifugation for 5 minutes at room temperature and then washed with a saline solution containing 5 % human albumin (FLEXBUMIN, Baxter). After washing, cells were frozen in CS10 medium (Stemcell) and stored at -150℃.
A GMP-grade lentivirus (produced by WuXi AppTec.) encoding the HBsAg-specific TCR gene was added to activated T cells on day 1. Cells were cultured at 37 °C for 8 to 12 days in a closed bag system (Cytiva) on a Xuri Cell Expansion System W25 (Cytiva). Cells were harvested by 300xg centrifugation for 5 minutes at room temperature and then washed with a saline solution containing 5 % human albumin (FLEXBUMIN, Baxter). After washing, cells were frozen in CS10 medium (Stemcell) and stored at -150℃.
4
Bacterial Adhesion on Tissue Grafts
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The BP patch (Supple Peri-Guard Pericardium; Synovis Surgical Innovations, St. Paul, Minn), CH tissue processed by the European Homograft Bank (EHB; stored at À80 C until use), and heterologous BJV (Contegra conduit; Medtronic, Minneapolis, Minn) were used to investigate bacterial adhesion. For the BJV conduit and CH, both the wall and valvular leaflets were used. The BP patch and BJV conduit were purchased from the manufacturers. The CH thawing process was done in accordance with instructions from the EHB. 7 All tissues were rinsed using 0.9% NaCl before use. A 10-mm Acu-Punch (Acuderm, Fort Lauderdale, Fla) was used to cut circular tissue pieces, as illustrated in Video 1. Pieces of BJV conduit were incubated overnight at 4 C with 200 g/L of human albumin (Flexbumin; Baxter, Westlake Village, Calif) to neutralize glutaraldehyde (protocol optimized during preliminary experiments), and were then rinsed with 0.9% NaCl. Bacterial adhesion was assessed for at least 5 to 17 different tissues pieces, except for the BJV leaflet (3 pieces). A graft tissue sample incubated in PBS served as a negative control.
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