The fist stage consists of inserting a fistula needle in a peripheral line. Should vene-puncture in the peripheral line not be possible, a central venous pressure (CVP) line in the format of an inner jugular line, a sub-clavian line or a femoral line, depending on the patient’s age, size condition and indication, is inserted into the patient using local anesthesia.

The second stage is the apheresis process. The machine we use is a Gambro COBE Spectra Version 7.0 made by Gambro BCT in USA (Denver, Co). This is continuous flow cell separator that splits the patient’s blood into 3 parts, RBCs, WBCs and Platelets. The RBCs and Platelets are returned immediately to the patient via the canella in the cubital vein (return line) while a small portion of the WBCs are taken from the patient. The total line fill in the apheresis kit is 161 ml which is entirely flushed back into the patient at end of the apheresis process. The diluents used are Saline and the anticoagulant used is ACDA solution made by Haemonetics and/or Baxter. This small portion of the WBCs apheresed from the patient (depends on the patient’s sex, height, weight, age, size and conditions as defined by the patient’s haematocrit reading), is the raw material for our therapy. The machine and process of apheresis is FDA approved and done regularly all over the world to collect RBC’s, WBC’s, Platelets and Peripheral Blood Stem Cells (PBSCs).

A highly purified CR3/43 murrain monoclonal antibody based on (New Zealand) calf serum is then added to the WBCs. This monoclonal antibody is raised against homologous region of the Beta chain of MHC class II antigens. This is then incubated at a steady temperature for a duration of between 3 and 7 hours in a tissue culture incubator. The cells are then removed from the incubator and saline washed for a duration of 20 minutes using a COBE 2991 Cell Processor made by Gambro BCT in USA. The centrifuge based washing allows for all additives to the cells to be washed out into the supernatant bag, which is then discarded. Cell washing is common practice in the haematology departments of all large hospitals worldwide.

The remaining cells, which are pure patient stem cells in a more committed state (progenitor cells), are then re-infused back into the patient via a normal drip set which is attached to the CVP line previously inserted. The patients is monitored for a period of between 24 and 48 hours to observe any reaction due to cell infusion and then discharged. There has been no reaction to the cell infusion in any of our patients to-date.

Murrain mono-clonal anti-bodies based on calf serum, similar to the ones used in the TriStem Retrodifferentiated Stem Cell Therapy are FDA approved for use in Baxter’s Isolex machine used to collect bone marrow stem cells. The Miltenyi machine approved by EMEA in Europe uses iron beads for the same extraction process. The monoclonal antibody binds with the stem cells to allow the machine to wash / extracting these particular cells.

TriStem Corporation’s CR3/43 murrain monoclonal antibody is exclusively manufactured for them by Dako Cytomation. This monoclonal antibody used by TriStem, although very similar to the ones made by BD and others that are used in the Baxter Isolex machine which are FDA Approved, is not FDA Approved as yet.