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 components
from the IRC
• Store blood components
• Pre-transfusion testing
• Patient’s ABO & Rhesus blood grouping
• Confirmation of donor’s ABO & Rhesus blood
group
• Crossmatching of donor’s & patient’s blood
• Issuing the compatible blood components to the
patient
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Apheresis services
• Apheresis for donor’s 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 cells
according to doctor’s 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 apheresis
2. 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 procedure
• Allogeneic
• 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 the
mobilization and leukapheresis procedure
• Non reactive for transfusion transmitted
infections (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 peripheral
blood – prior to collection
• Using hematopoietic growth factors G-CSF
5 – 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 PBSCs
collection
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CD34+ cells pre-count

• WBC count alone does not predict the CD34+
cells yield
• Peripheral blood CD34+ cell count corralates
with the final cell yield
• No correlation between the pre-apheresis
CD34+ cell number and the percent CD34+
efficiency
• It is well known that the number of cycles of
chemotherapy / inversely affects the collection
efficiency and yields of CD34+ cells
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Collecting the stem cells

• Day 5 is an appropriate time to start apheresis
• If not sufficient, the second apheresis on day 6
could be performed (G-CSF still given to patient
on day 5 )
• A minimal of pre-apheresis CD34+ cells
concentration 30 / ul can optimize yield and
maximize cost effectiveness
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Collecting the stem cells

• Peripheral blood CD34+ cells concentration
exceeded 40 / ul more than 2.0 x 106 CD34+/ kg
BW could be expected by a single apheresis
• Absolute efficiency in allogeneic PBSCs donors:
35 % - 45.5 %
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Collecting the stem cells

• Prophylactic oral calcium given prior to
apheresis procedure
• Collection performed via peripheral or central
venous
• Using apheresis machine:
• Continuous flow centrifugation, or
• Intermittent flow centrifugation
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Continuous flow centrifugation

Cobe Spectra
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Intermittent flow centrifugation

MCS+
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Targets for stem cells collection

• 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 febrile
complications 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)
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Preservation of hematopoietic stem

cells
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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 ficoll
density separation and washing procedure
• Needs peristaltic pump (40 ml / minute) to add
the 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 in
biological 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 ° C
for at least 15 min
• Slowly mixing the same volume of stem cells and
cryoprotectant
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Cryopreservation
• 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 )
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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