A Validated and Licensed Method of Bone Marrow Aspirate Procurement to Support Advanced Therapy Investigational Medicinal Product (ATIMP) Manufacture

The Blood & Tissue Establishment (BTE) of Galway University Hospitals supplies sterile bone marrow (BM) aspirate to European cellular manufacturing facilities as a starting material in ATIMP manufacture. We have developed a validated, sterile bone marrow procurement method in accordance with the European Union Tissue & Cells Directive (EUTCD) and Good Manufacturing Practice (GMP). The BTE achieved the requisite procurement licence, from the Irish competent authority, the Health Products Regulatory Authority (HPRA).

Under the BTE Quality Management System all standard operating procedures and forms are document controlled, all consumable items are batch accepted, all staff involved undergo training and competency assessment (including adequate sanitisation of the iliac crest), and all issues are recorded and reported appropriately to the HPRA.

The prospective donor must first undergo a two part 'donor assessment' procedure which evaluates the donor's 'fitness' to undergo the procedure, while also having due regard for any potential recipient.

Part One of the assessment process requires that the entire procedure is explained by a trained clinician / nurse; a patient information leaflet is provided and written consent obtained. A full blood count, coagulation screen, glucose test, chemistry screen, blood group and virology screen is performed. The virology screen must, at minimum, include the viruses listed by the EUTCD and the donation must take place within 30 days of sample collection. The patient will then undergo a 'lifestyle questionnaire' to assess donor medical, travel, sexual and other risk history. Guidance for allowing the donation to proceed will depend on a number of factors, including, whether it is an allogeneic or autologous donation. Consideration is given to the 'Bone Marrow and Peripheral Blood Stem Cell Donor Selection Guidelines for Unrelated Donors' of the 'Guidelines for the Blood Transfusion Services in the United Kingdom'. HPRA and European Centre for Disease Prevention and Control alerts are also incorporated into selection criteria.

The Responsible Person (RP) of the BTE (also a Consultant Haematologist) must review the answers to the questionnaire, together with the blood test results and assess whether the donor is fit to proceed to aspiration.

Part Two of the assessment process takes place immediately prior to the scheduled aspiration. The donor will repeat the lifestyle questionnaire once more and the answers will be reviewed and compared with the previous questionnaire. The Consultant Haematologist will carry out a clinical examination and assessment of the donor. Upon completion, final donor eligibility is signed off and the aspiration can proceed. The procedure takes place in an operating theatre by a scrubbed Consultant Haematologist. A sterile field is created around the aspiration site. The aspiration site is cleansed with Chlorhexadine. Local anaesthetic (lidocaine 2%) is administered. 11 Gauge BM needles are used to puncture. Syringes are flushed with heparin sodium flushing solution. The flushed syringes are used to withdraw the BM, handed to a sterile assistant and transferred into 10ml lithium heparin vacutainers. The volume of 30ml of BM is aspirated from the iliac crest of both healthy donors (allogeneic) and patients (autologous) enrolled in clinical trials.

These vacutainers are capped and inverted several times to prevent clotting, labelled and packaged for transport.

The puncture site is dressed, donor vitals are observed and noted post procedure and the donor may be discharged after 2 hours.

BM is transported to the manufacturing facility at monitored ambient temperature. Within the Grade A controlled environment, the BM is pooled and 1ml is used to inoculate one aerobic and one anaerobic blood culture bottle (validated for 0.5ml in each, incubated for 14 days, automated detection system).

As of July 2017, we have performed 24 collections, 11 of which are allogeneic donations from healthy volunteers. The average white cell count in the aspirate from healthy donors is 43.1 x 10⁶/ml. The sterility is satisfactory, with only one instance of contamination over the 24 collections, where, on the final day of incubation (day 14), we detected propionibacterium spp, a notoriously slow-growing commensal skin flora.

Our regulatory approved protocol provides a steady supply of GMP grade bone marrow aspirates for the purpose of clinical trials.