Human Blood-Derived Raw Material:
Enabling Controlled, Consistent Collection

 

 

 

Thomas V. Ramos, MS, MBA, William F. Busby, III, Gaytha McPherson

HemaCare Corporation

(877) 397-3087 • tramos@hemacare.com

 

Scott R. Burger, MD

Advanced Cell & Gene Therapy, LLC

(919) 969-1103 | celltherapy@ac-gt.com

 

 

Abstract

Human cells are critical raw materials for manufacturing cell therapy products, but often introduce significant variability. Rigorous operational controls and quality systems, however, enable optimal collection of high-quality, consistent cellular material. HemaCare, a long-standing supplier of human-derived blood components, controls apheresis procedures and collection sites under a formal quality system, with GMP-compliant, validated procedures and equipment, and GTP-compliant donor screening and tracking.

HemaCare performed 69,658 cellular apheresis collections in the last five years, including patient and normal-donor PBMCs, G-CSF-mobilized PBPCs and plateletpheresis products, for research, clinical trials, and commercial products.

Expanded capabilities include disease-state and normal-donor bone marrow, umbilical cord blood, and cord tissue collection, immunomagnetic cell selection, cryopreservation, and analysis by flow cytometry.

HemaCare unmobilized apheresis products showed consistently high MNC purity, with 93.8% of products containing ≥ 75% MNC, and an average of 85.2% MNC ± 6.6% (mean ± 1 SD).

Red blood cell contamination was low, with hematocrit averaging 1.8% ± 0.8%.

Approximately 85% of HemaCare donors have donated apheresis products 5 or more times, and this repeat-donor pool also contributes to product consistency, as MNC content of individual donor apheresis products had an average coefficient of variation of 3.5%, compared to a CV of 7.7% for all apheresis products.

 

About HemaCare

HemaCare is a leading provider of apheresis products, human blood cells, apheresis collection services, and therapeutic apheresis services

• Apheresis collections and blood-derived products for preclinical research, clinical studies from Phase I to Phase IV, and commercial applications
• Supports applications in immunotherapy, cell therapy, assay development, and medical devices

• Apheresis PBMC
• G-CSF-mobilized PBSC
• Bone marrow
• Cord blood
• Peripheral blood
• Plasma, serum

• Cell subpopulations
  • CD34⁺
  • CD3⁺, CD4⁺, CD8⁺
  • CD19⁺, CD56⁺, others
• Healthy-donor and disease-state products
• Fresh and cryopreserved products

 

The HemaCare Advantage

HemaCare is committed to providing our customers with experienced, personalized, responsive, cost effective, and value added services.

 

Research Products and Cellular Therapy Services

• Donor pool is already pedigreed and will continue to be expanded
• Extensive donor registry with ability to request repeat donor collections
• Predictable, reliable, and validated collection procedures
• Optimized Standard Operating Procedures leading to high degree of standardization and control
• Ability to collect based on specific, customizable protocols
• High-yield, consistent cell collections
• Validated, automated cell counts and five-part WBC differentials
• Established distribution redundancies leading to the ability to ship via FedEx, UPS, World Courier, and various local couriers
• Access to our scientific/technical support 24/7/365

Human Cells: Standardizing Living Biological Raw Material Through Quality Processes

• Human blood-derived cells are critical raw material for cell therapy, tissue-engineered products, and ex vivo gene therapy products
• Quality and consistency of cellular raw material is a major determinant of final product characteristics
• Controlling cell collection minimizes variability and increases likelihood of success in research and manufacturing
• Training and experience are critical
• Quality Systems standardize and control operations

Controlled collection procedures yield optimal, consistent products

 

Quality Indicators

• Tracking and trending
  • Donor reactions, deviations, exceptions, suppliers, risks, equipment performance, etc…
• Product QC analysis
  • Automated cell counter/analyzer, 5-part WBC differential (Horiba Pentra analyzer)
    • Nucleated cell (WBC) content and subpopulations, % MNC, Hct, product volume, etc…
  • Immunophenotyping (MACS Quant flow cytometer)
• Donor testing
  • Screening, infectious disease testing, CBC with 5-part WBC differential
• Internal and external audits

 

Selected Apheresis Product Quality Indicators
MNC Purity/Content, RBC Contamination

MNC Purity 85.2% ± 6.6% (mean ± 1 SD) 93.8% of products ≥ 75% MNC   MNC Content 10.6 ± 3.8 x 109 MNC (mean ± 1 SD)         RBC Contamination Hematocrit 1.8% ± 0.8% (mean ± 1 SD) 91.3% of products £ 2.5% hematocrit

 

Donors – The Critical Source

• All donors are qualified per regulations and protocol requirements, with IRB-approved informed consent
• Pedigreed, well-characterized apheresis donor population
  • 85% of HemaCare donors have donated ≥ 5 times/year
  • Facilitates recruitment of donors with specific characteristics required by investigator
    • Medical history, HLA type, other laboratory test results, age, gender, ethnicity…
  • Repeat donors further minimize variability

% MNC Mean 3.5% CV for products per donor Mean 7.7% CV for all products n=21 donors, 3-5 products/donor

 

External Quality Indicators
Dendreon Supplier Scorecard

External Quality Indicators

Non-Cryopreserved PBMC Storage

• For many applications, PBMC apheresis products must be used or cryopreserved within 24-48 hours post-collection. Storage in HypoThermosol^® (BioLife Solutions) has been shown to increase stability of a variety of cell types, and could extend shelf-life of apheresis PBMCs.
• HemaCare has begun testing stability of MNCs and cell subpopulations stored in HypoThermosol for up to 96 hr without cryopreservation.

 

Test at t = 0, 24 hr, 48 hr, 96 hr   Cell count with differential Total WBC and MNC content % Viable cells (7-AAD) % Viable MNC recovery   Viable CD4+, CD8+, CD14+, CD19+ cell frequency

 

Non-Cryopreserved PBMC Storage
Plot Data

Summary

• Collecting blood-based cellular products in a manner that minimizes variability brings a higher degree of reproducibility to the research project or manufacturing effort
• Quality-based controls such as standardized SOPs, staff training and competency assessments, equipment management, and monitoring of quality indicators reduce this variability
• Availability of repeat donors from a pedigreed donor base enhances the quality and value of this critical, living biological material
• Use of cGMP, serum-free, protein-free biopreservation media such as HypoThermosol shows great promise to enable worldwide shipment of fresh cellular products isolated from apheresis collection, extending shelf-life of cell therapy products, and delaying need for cryopreservation