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8/8/2019 Role of Blood Bank in Stem Cells Therapy
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Role of blood bank in stem cells
therapy
Auda S. Aziz
Dharmais Cancer Hospital
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Topics
Blood bank activities
Collection of stem cells by apheresis Stem cells preservation
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Dharmais Blood bank activities
3 groups of activities:
Blood component services
Apheresis services
Stem cells preservation
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Blood component services
Receive stock of screened-blood componentsfrom the IRC
Store blood components Pre-transfusion testing
Patients ABO & Rhesus blood grouping Confirmation of donors ABO & Rhesus blood
group Crossmatching of donors & patients blood Issuing the compatible blood components to the
patient
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Apheresis services
Apheresis for donors blood component Screening the blood donor Performing apheresis for required blood
component, esp. Platelet Concentrate
Therapeutic Apheresis Screening the patients
Performing thrombapheresis, leukapheresis,plasmapheresis or apheresis for red cellsaccording to doctors requirement
Apheresis for stem cells (PBSCs)
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Stem cells preservation
Collection of MNCs
Cryopreservation of MNCs Thawing the MNCs before infusion
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Role of blood bank in stem cells therapy
1. Collects the hematopoietic peripheral blood
stem cells (PBSCs) by apheresis2. Preservation of hematopoietic stem cells:
Bone marrow derived hematopoietic stem cells
Peripheral blood hematopoietic stem cells
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PBSCs collection by apheresis
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Eligibility of patients / donors
Autologous
Patients eligibility for the apheresis procedureAllogeneic
Donors physical ability for the procedures
The willingness of the donors
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Patients / donors selection Selection criteria
Minimal Hb level : 8 g / dl
Platelet count > 150,000 / ul
Acceptable conditions to undergo themobilization and leukapheresis procedure
Non reactive for transfusion transmittedinfections (for allogeneic transplant )
Acceptable CD34+ pre-count
Informed consent
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Mobilizing the stem cells
Mobilizing the hematopoietic stem cells from
marrow compartment into the peripheralblood prior to collection
Using hematopoietic growth factors G-CSF5 10 ug / kg subcutaneus, for 5 days
For malignant blood diseases : G-CSF 5 10 ug / kg with chemotherapy
G-CSF alone has a poor yield in PBSCscollection
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CD34+ cells pre-count
WBC count alone does not predict the CD34+cells yield
Peripheral blood CD34+ cell count corralateswith the final cell yield
No correlation between the pre-apheresisCD34+ cell number and the percent CD34+
efficiency It is well known that the number of cycles of
chemotherapy / inversely affects the collectionefficiency and yields of CD34+ cells
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Day 5 is an appropriate time to start apheresis
If not sufficient, the second apheresis on day 6could be performed (G-CSF still given to patienton day 5 )
A minimal of pre-apheresis CD34+ cells
concentration 30 / ul can optimize yield andmaximize cost effectiveness
Collecting the stem cells
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Peripheral blood CD34+ cells concentration
exceeded 40 / ul more than 2.0 x 106
CD34+/ kgBW could be expected by a single apheresis
Absolute efficiency in allogeneic PBSCs donors:35 % - 45.5 %
Collecting the stem cells
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Prophylactic oral calcium given prior toapheresis procedure
Collection performed via peripheral or centralvenous
Using apheresis machine:
Continuous flow centrifugation, or
Intermittent flow centrifugation
Collecting the stem cells
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Continuous flow centrifugation
Cobe Spectra
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Intermittent flow centrifugation
MCS+
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For autologous transplant
Minimal threshold of CD34+ : 0.75 1 x 106 / kg
Recent data support an economic benefit associated
with greater CD34+ cell collections : 5.0 x 106 / kg (accelerated platelet engraftment, reduced febrilecomplications and use of antibiotics after transplant )
For allogeneic transplant
Minimum dose of 2.0 x 106 / kg of CD34+ cells For stem cells therapy
Minimal dose of 25 x 106 of CD34+ cells (AMI)
Targets for stem cells collection
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Preservation of hematopoietic stemcells
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Cells selection
Collection of MNCs by adding the ficol solution
Using cobe 2991 ( for stem cells transplant ) Tube method ( for stem cells therapy )
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Cobe 2991 Is the automated cell washing machine
Used manually to isolate the BM-MNCs by ficolldensity separation and washing procedure
Needs peristaltic pump (40 ml / minute) to addthe BM into the spinning ficoll solution
Input 550 ml, output 100 ml
Ficol + MNCs
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Tube method The manually isolation of BM-MNCs by ficoll
density separation and washing procedure
Using sterile consummables, performed inbiological safety cabinet
The volume of BM processed 50 ml
The final product volume 4 10 ml
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Cryopreservation Cryoprotectant preparation :
20 % DMSO
20 % autologus plasma In normal saline
Incubate stem cells and cryoprotectant at 4 Cfor at least 15 min
Slowly mixing the same volume of stem cells andcryoprotectant
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Controlled rate freezing
Minus 1 C / min from 4 C to minus 40 C
Minus 5 C / min to minus 160 C Store in liquid nitrogen ( minus 196 C )
Cryopreservation
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Evaluation
Cell count
Total nucleated cell concentration
Mononuclear cell concentration CD34+ cells , etc
Sterility test (aerobic, anaerobic)
Cells culture
Samples : Starting materials
Final product
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Thawing
Perform near patient ward
Immediately infused to the patient
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Thank you
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