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Once you decide to bank you baby's cord blood stem cells, your bank will send you a collection kit with instructions on what to do next. Collection kit contents vary from bank to bank but there are some standard items that all kits should contain. Your collection kit should contain the following:

• Directions for you, on what to do with the kit before and after your delivery.
• Directions for your doctor.
• A container with packing material and instructions.
• Labels to uniquely identify your baby's cord blood.
• A blood collection bag.
• A sterilized insertion needle to allow the blood to flow into the collection
  bag(s).
• Blood collection tubing to connect the inserted needle to the collection bag.
• Umbilical cord prep kit to prepare and sterilize the umbilical cord before the needle is inserted.
• Clamps so that the blood collection tubing and umbilical cord can be clamped when required.
• Maternal blood tubes to collect the mother's blood.

On your way to the hospital, inform your bank that you are about to deliver. Most, if not all banks can be reached by phone 24 hours a day, 7 days a week. Your bank will then arrange for a medical courier (who should be available 24 hours a day) to pick up the sample and deliver it to the lab within 48 hours.

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Collection of cord blood is an essential part of cord blood stem cell banking. There is only a 15-minute time frame to collect the cord blood. Any delay beyond the stipulated time will result in less cord blood being collected and therefore fewer stem cells. The collection kit must be kept handy and you must ensure that your doctor is aware of your decision to bank your baby's cord blood and willing to carry out the procedure.

The collection process occurs after your baby is born and the umbilical cord is clamped and cut. The blood can be collected before or after the delivery of the placenta.The two common methods of cord blood collection are:

• In Utero- here the cord blood is collected after the baby is delivered but the placenta is still in the uterus. This is an invasive procedure and must be done with full consideration of any adverse effects on mother and child even though the risk of contamination is lesser.
• Ex Utero- In this case the cord blood is collected after the baby and the placenta are delivered. For cesarean births, the obstetrician's focus is on the surgical needs of the mother and this method is especially recommended so that the cord blood can be collected separately.

  If the collection is ex-utero, then the placenta is placed in a sterile container with the umbilical cord hanging down. Again, there are two ways in which the blood can be removed from the umbilical cord. A needle can be inserted into the umbilical cord and the blood allowed to drip by gravity into a 'bag' or be drawn out with a 'syringe.' The bag method limits exposure to airborne bacteria and reduces the risk of contamination, as it is a 'closed' system. Most doctors also prefer to let the blood drain by gravity into a bag.

Once the blood is collected, the bag is sealed carefully to avoid contamination. Your cord blood bank should provide specific instructions regarding the sealing and packing of the collection kit so that there is no room for error. The sample should be packed securely in a bag that can be sealed and then packed in the box.

Once the blood is collected, the bag is sealed carefully to avoid contamination. Your cord blood bank should provide specific instructions regarding the sealing and packing of the collection kit so that there is no room for error. The sample should be packed securely in a bag that can be sealed and then packed in the box.

Cord blood should be maintained at room temperature till it reaches the lab because a sudden change in temperature may destroy the cells. Room temperatures vary from country to country and so some companies, depending on their location, may provide a frozen gel pack in the collection kit. This gel pack helps maintain the temperature of cord blood.

It is also extremely important to label the bag correctly. Ideally, your bank should generate a unique bar coded label and send it with the collection kit so that the label can be applied to the sample after collection. This makes it easy to identify and track the cord blood sample once it leaves the hospital.

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To maintain cell viability, the timing, pressure and temperature regulation are of extreme importance, all of which are controlled by the use of a private medical courier. This ensures that the cord blood unit is kept at the proper temperature and pressure during transport and delivered to the laboratory within 48 hours. This time regulation will help maximize the number of cells that can be used in a transplant should you ever need them.

On your way to the hospital, inform your bank that you are about to deliver. Your bank will then arrange for a medical courier (who should be available 24 hours a day) to pick up the sample and deliver it to the lab within 48 hours.

The kit is shipped once the cord blood has been collected, safely sealed and packed.

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Once the cord blood unit reaches the lab, samples from the unit are sent for bacterial culture and blood grouping. The maternal blood is tested for transmissible diseases such as HIV and Leptospirosis. If the sample tests positive, then the cord blood is also tested to see if the disease has been passed on. Depending on the outcome of the tests, the cord blood sample may be quarantined or rejected. The tests that are carried out on the maternal blood and cord blood, and the fate of a sample that tested positive for a disease is determined by government guidelines for the country in which the bank is located.

After testing, the remaining cord blood undergoes processing in two stages volume-reduction and separation to isolate stem cells.

Volume reduction can be accomplished either by sedimentation or spinning in a centrifuge. To process the cord blood, a chemical called "Hespan Starch" or "Hetastarch" is added. Hespan solution facilitates the separation of red cells from the mononuclear white cells during sedimentation and centrifugation. The blood components are separated so that: red cells are at the bottom, plasma (a clear white liquid) is on top, and in the middle is a pinkish layer called the "buffy coat" which contains the white blood cells (WBC), including stem cells. At this point, a gentle spin in a centrifuge is sufficient to isolate the white blood cells (WBC).

Separation of stem cells is accomplished by further centrifuge spinning of the white cells.

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The array of tests performed on the maternal blood help in deciding whether the cord blood sample will be stored or not. A blood sample that tests positive for HIV may mean that the virus is present in the cord blood sample. Many governments including the Indian government do not allow blood banks to store HIV positive samples. All this should be communicated to the parents. A report of all testing done along with results must be sent to parents for their information and future reference. The report will also document the fact that these tests were actually performed by the lab.

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After the cells have been separated, the unit containing stem cells is mixed with a cryopreservation solvent. This solvent protects the cells against the formation of ice crystals, which can rupture the cell membrane. The solvent most commonly used for cryopreservation of cord blood is "DMSO" (Dimethyl Sulfoxide, usually in 10% solution).

To further protect cord blood cells against the formation of ice crystals, it is important that the cells freeze slowly. This is done in a special controlled-rate freezer, and then the blood is transferred to long-term storage. The final storage temperature should remain stable at -196^o C as temperature fluctuations will cause the blood cells to suffer irreparable damage.

The samples are stored in bags, in insulated freezers cooled by liquid nitrogen or vapor nitrogen. The advantage of liquid nitrogen is that the temperature maintained is constant and not subject to fluctuations as is the case with vapor nitrogen. However, you can never be too safe where your child's cord blood is concerned and so freezer temperatures must be monitored at all times as an extra precaution.

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Stem cells and cord blood should be processed in a clean room i.e. a room with only clean, filtered air - if there is no closed system for processing the stem cells. The air entering a clean room from outside is filtered to exclude dust. In a clean room, the air inside is constantly re-circulated through High Efficiency Particulate Air (HEPA) and Ultra Low Penetration Air (ULPA) filters to remove contaminants. The clean room facility should be a Class 100 facility.

Clean room sterility is further maintained by staff who enter and leave through airlocks and wear a clean room suit i.e. protective clothing such as hats, face masks, gloves, boots and cover-alls.

The filtration process ensures that there is no chance of contamination from the environment. The clean room suit also known as a 'bunny suit', prevents any hair and skin from being shed in the laboratory. The suit also prevents microbes that may be present on the skin from coming in contact with samples in the laboratory. These precautions ensure that the cord blood and stem cells do not get contaminated. Contamination by dust particles reduces cell viability whereas, microbial contamination can infect the samples and destroy the cells within.

The lab must have a back up generator that kicks in anytime there is a power shortage or voltage fluctuation. There should also be adequate measures like fire extinguishers and automatic sprinklers in the event of a fire. Apart from this, the lab should be monitored 24 hours a day and 7 days a week to manage any problem such as equipment failure, accidents etc. All automated processes, freezer temperatures and laboratory temperatures should be monitored at all times, as they are vital for the preservation of stem cells.