Procurement of fetal precursor cell transplants
Procurement of fetal precursor cell transplants must be carried out under aseptic conditions in the manufacturing laboratory, following the GMP and GLP rules, with a complete documentation.
The crucial link between the ‘scheduled’ rabbit females from the closed colony as sources of fetal and newborn and rabbits used in the procurment of fetal precursor cell transplants, and a patient/recipient of the fetal precursor cell transplants necessary for the treatment, identified by a code, must not interrupted throughout the entire production cycle. It is accomplished in the following steps of manufacturing of fetal precursor cell transplants by BCRO method:
1/ All fetal precursor cell transplants prepared for one specific named patient for a predetermined date of implantation are classified as a ‘batch’.
2/ All fetal precursor cell transplants in a batch are prepared from fetuses and newborns of ‘pre-scheduled’ rabbit females only.
3/ Each sterilized box (for transportation between rabbit colony and manufacturing laboratory) contains only one ‘pre-scheduled’ rabbit female, to be used as source of fetuses, marked by identification number of that rabbit female. Or, each sterilized ‘small’ transportation box contains only newborns of one pre-scheduled rabbit female, marked by identification number of the rabbit female. All ‘small’ boxes with rabbit newborns necessary for manufacturing of a batch of fetal precursor cell transplants for one named patient are placed inside of one large card box.
4/ All ‘source’ animals are processed in groups. One ‘group’ equals all fetuses of one rabbit female or all newborn rabbits from one small box, i.e. from one female.
5/ All euthanized animals of one group are placed in a metal container marked with an identification number of a rabbit female, the ‘source’ of all fetal or newborn cadavers in the metal container.
6/ Veterinary doctor/ pathologist carries out a full autopsy, properly evaluates gross findings, and collects specimens for histological and microbiological testing, if necessary. In case of any pathological findings the source animal is discarded. These findings are placed in the permanent record of each fetal precursor cell transplant, that contains also the summary of medical records of all rabbit females whose fetuses or newborns are used in the preparation of the batch of fetal precursor cell transplants for a specific patient.
If findings are normal during the autopsy, the veterinary doctor dissects all organ(s) and tissue(s) needed for the production of all fetal precursor cell transplants of a batch, collects them in the Petri dishes, labeled with
- the name of fetal precursor cell transplant scheduled to be produced,
- the sex of the recipient / patient that must be the same as the sex of all animal sources used for the manufacturing of fetal precursor cell transplants of placenta and adrenal cortex,
- the identification number(s) of the rabbit female(s), and
- the code of the patient / recipient of the batch of fetal precursor cell transplants under preparation.
7/ After mechanical mincing by scalpel or scissors, the pre-determined quantity of tissue fragments of each organ or tissue is placed in the tissue culture flask, labelled with the name of fetal precursor cell transplant, identification numbers of all rabbit females whose fetuses and newborns have been used in preparing the tissue culture in the respective culture flask, and a patient's code.
After the final step of manufacturing process, collection of cultured tissue fragments into transportation vials ,
8/ each transportation vial contains the individualized treatment dose of one fetal precursor cell transplant of a batch, properly labeled (see below in chapter on ‘labeling’), including an identification number of fetal precursor cell transplant, which number is linked to all fetal and newborn rabbits used in the manufacturing of a fetal precursor cell transplant placed in that vial via the identification numbers of all females, ‘sources’ of the same fetuses or newborns,
as well as linked to the
- detailed records of entire manufacture,
- histological verification of the organ or tissue origin of each fetal precursor cell transplant,
- records of microbiological testing,
- records of bacterial endotoxin test,
- records of other tests for quality control and validation.
On the last day of manufacture of a batch of fetal precursor cell transplants for a named patient
three samples of supernatant of each stem cell transplant are taken as well
1st sample is kept by manufacturer in liquid nitrogen for 5 years:
2nd sample is used for microbiological testing;
3rd sample is used for bacterial endotoxin test.