Cellgenea^TM

Cellgenea - the innovative service of the Pharmaceutical Biotechnology Company Biogenea-Cellgenea Ltd - focuses on the isolation of the cord blood progenitor hematopoietic cells, for cryopreservation and the possible future use, after ex-vivo cell expansion using the super automated bioreactors of Regenetech and the technology obtained from NASA.

The procedure is simple, is carried out at the end of the delivery from specialized personnel and it is completely painless and hazard-free both for the neonatal and the mother. The procedure consists of the following steps:

Collection

Cord blood is being collected from the cord vessels immediately after the delivery and after it’s cut from the neonatal. Normally this blood is thrown away, so the collection doesn’t affect at all the delivery procedure. The collection is carried out by specialized personnel and the procedure is the same either in normal delivery or in cesarean section.

Special guidelines are enclosed for the personnel for the cord blood collection, in order to collect the maximum quantity of blood avoiding the possibility of any contamination.

The cord blood is delivered in a special collection KIT that protects it from any possible irradiation, mechanical damages, and furthermore in stable temperature using thermo stable gel packs.

In utero or ex utero cord blood collection: which is better? Lasky LC et al. Transfusion. 2002; 42:1261–1267

Sterile processing and progenitor cells isolation

The sample is being given a unique barcode for its traceability both during the processing and after the cryopreservation.

Sample processing is being carried out immediately upon receipt, with validated procedures in a completely closed system (Pall® cryoset). In particular, the total nucleated cells (cells with a nucleus, like the progenitor Hematopoietic cells) are being isolated form the red blood cells and the plasma through sedimentation and centrifugation. The whole procedure is being carried out in cleanroon class A/B (Annex I - February 2008 EU guideline for the cGMP standards). There is a complete monitoring system inside the cleanrooms 24h a day controlling the air particles as well as the temperature and the humidity. In addition, the surfaces (laboratory and personnel) are controlled periodically for their germ load (there is a standard range for both germ and air particles inside the cleanrooms). The scientific personnel are properly trained for the procedure, so the whole procedure is being carried out in sterile conditions.

A Rapid Closed System for Isolation of Mononuclear Cells From Whole Blood Based on Pall Leukocyte Reduction Technology, L Bradbury et al.

Quality Control

Cell counting

A cell count is being performed both prior to and after the procedure. Prior to the procedure a full blood test is being performed, and the white blood cells, as well as their subgroup are being counted. The blood analysis is carried out using the automated blood analyzer AcT-5 dif (Beckman Coulter).

After the procedure completion, a small amount of the final product is being used for the final numerical counting of the progenitor cells, as well as their viability prior to the cryopreservation using flow cytometry using the ISHAGE protocol.

The ISHAGE guidelines for CD34+ cell determination by flow cytometry. International Society of Hematotherapy and Graft Engineering. Sutherland DR, Anderson L, Keeney M, Nayar R, Chin-Yee I. J Hematother. 1996 Jun;5(3):213-26.

Further tests after the cryopreservation

A small amount of the sample is being cryopreserved and thawed especially for a viability test of the progenitor Hematopoietic cells after  the cryopreservation procedure. Using this procedure we can guarantee that the viability and the plasticity of the progenitor cells remain intact during and after the cryopreservation. The viability test is being performed using the automated luminometric system Glomax of Promega (the first one installed in Greece), while the plasticity test is being performed using the Methocult® system of StemCell Technologies.

Adherent cells generated during long-term culture of human umbilical cord blood CD34⁺ cells have characteristics of endothelial cells and beneficial effect on cord blood ex vivo expansion. Yoo E et.al., Stem Cells 21: 228-235, 2003

A small amount of the plasma si controlled for the presence of aerobic and anaerobic germs using the automated  blood culture system BacT/ΑLERT BioMérieux, while in parallel the new automated immunological analyzer Architect i1000SR (Abbott Diagnostics) (the first one installed in Greece) for the detections of HBsAg, as well as antibodies for Hepatitis C (Anti-HCV), HIV Ι/ΙΙ (Anti-HIV I/II), Toxoplasma (Toxo IgM), Syphilis and CMV IgM. 

Evaluation of Assay and Workflow Performance of the ARCHITECT^® i1000SR Immunoassay Analyzer Hong al., 2007 American Association for Clinical Chemistry Annual Meeting, San Diego, California, July 15-19, 2007

Incubation time required for neonatal blood cultures to become positive. Jardine L et al. J Paediatr Child Health. 2006, 42(12):797-802.

Ex-vivo cell expansion

Cellgenea in exclusive collaboration with Regenetech uses patented technology from the research laboratories of NASA for the expansion of progenitor cells .

As a result, the isolated cells could be expanded in case of a future use.

Cryopreservation and Release of the Product

The isolated progenitor Hematopoietic cells are stored in temperatures reaching -196⁰C. The results are sent along with the certificate of cryopreservation only after the completion of all the quality tests. In these conditions all the metabolic procedures are stopped. Both the temperature, as well as the liquid nitrogen level are continuously controlled.

Cellgenea secures the immediate and validated cell release of the progenitor Hematopoietic cells in case of a future use.

Preservation of immunological and colony-forming capacities of long-term (15 years) cryopreserved cord blood cells. 28Koylka P et.al.,Transplantation. 1998;65:1275-1278.

Therapeutic applications

The continuous advance in the future applications of the progenitor cells for therapeutic use is overwhelming. Thus, the fact that these cellular populations are being used since many years for the treatment of autoimmune diseases as well as a supporting treatment for some cancers is being ignored. In particular, after specific treatments such as chemotherapy or irradiation (that destroy the immunological system) a blood system reconstruction is needed.

New applications of the progenitor cells are continuously emerging. Especially in the regenerative medicine, these cell populations are applied in order to heal tissues and organs. 

The cord blood progenitor hematopoietic cells have in general more potency for differentiation than the adult stem cells. There is a future prespective for the treatment of diabetes, brain damage, neuroral disorders, Parkinson's disease and multiple sclerosis. Furthermore, heart infraction and seizures could be added to the list of future treatments.

The treatment of many pathological conditions is already in preclinical phase. As a result, a life insurance by cryopreserving cord blood progenitor Hematopoietic cells could be a preventing tool for the contemporary medicine. 

30Umbilical cord blood research: current and future perspectives. Newcomb JD et al., Cell Transplant. 2007;16:151-158.

31Multilineage differentiation activity by cells isolated from umbilical cord blood: expression of bone, fat, and neural markers. Goodwin HS et al.,  Biol Blood Marrow Transplant. 2001;7:581-588.

32Umbilical cord blood-derived cells for tissue repair. Korbling M et al., Cytotherapy. 2005;7:258-261.

33Umbilical cord blood hematopoeitic stem cells and therapeutic angiogenesis. Goldberg JL et al., Cytotherapy. 2007;9:4-13.