Method for collecting fluids

A method for collecting blood or fluids in general from a placenta or organ or body tissue through the cut end portion of the umbilical cord or of one or more vascularization vessels or outflow vessels of an organ or tissue, including the steps of connecting the cord, or vessel, to a collection apparatus and then applying pressure to the placenta, or organ or tissue.

BACKGROUND OF THE INVENTION
 The present invention relates to a method for collecting blood or other
 fluids from an organ or tissue through a vessel. The method according to
 the invention is used in the biological, medical and veterinary field for
 diagnostic and therapeutic purposes and for research purposes to collect
 blood or other fluids from organs or parts of organs or tissues. In
 particular, the method according to the invention is used to collect
 placental blood, that is to say, the blood that is present in the
 placenta, during childbirth, directly after the birth of the neonate.
 In the biological, medical and veterinary fields it is necessary to collect
 fluids, particularly blood, from organs or tissues or containers in
 general, both for diagnostic and therapeutic purposes and for research
 purposes. Two non-limitative examples are: 1. the collection of blood from
 masses of tumoral tissue, since the blood contained in the tumoral tissue
 is potentially useful for therapeutic purposes after appropriate
 treatment; 2. the collection of blood from the placenta during childbirth
 directly after cutting the umbilical cord. In this second case, the blood
 is useful since it contains stem cells, which are hematopoietic precursor
 cells, that is to say, cells which are capable of reconstituting the
 hematopoietic system and therefore can be used, if the need arises, for
 transplanting to the same donor or to another compatible recipient.
 The procedures currently used to collect placental blood from an umbilical
 cord are described hereinafter as a typical example of the recovery of
 fluids from an organ; it is important to note that the reference to this
 practice is not limitative and is merely an example of situations in which
 it is necessary to collect blood or another fluid from tissues, organs or
 other containers in general.
 Placental blood from the umbilical cord is collected because it contains
 hematopoietic stem cells which can be used for transplants. To allow the
 collected blood and therefore the recovered cells to be truly and
 successfully usable, the following conditions must be met: 1. the largest
 possible amount of blood, that is to say, the highest possible number of
 cells, must be recovered; 2. the collected blood, which is fetal in
 origin, must not be contaminated by foreign cell populations, such as for
 example maternal cells; 3. during collection, contact of the blood with
 the outside (air or other potentially contaminated objects) must be
 minimized, or the collection procedure must occur in sterile conditions,
 possibly in a closed environment; 4. the possibility of human error must
 be minimized.
 Steps of Collection
 a. Before Expulsion of the Placenta
 Blood is currently extracted from the placenta in the medical field, both
 for diagnostic and therapeutic purposes and for research purposes, as
 follows: during childbirth, directly after the expulsion of the neonate,
 the umbilical cord is closed (clamped) in two points and cut in a point
 which is intermediate between the two closure points. After cutting, the
 cut end of the umbilical cord which is connected to the placenta protrudes
 freely from the mother's vagina and is available for collection
 operations. Typically, blood is collected in the period between the
 cutting of the umbilical cord and expulsion of the placenta. During this
 period, the flow of blood is ensured by the pressure generated by uterine
 contractions, which by compressing the placenta facilitate the outflow of
 blood through the umbilical cord.
 b. After Expulsion of the Placenta
 Only a small number of authors reports collection procedures which provide
 for cord blood collection after expulsion of the placenta by placing the
 placenta on a frame and collecting the blood by gravity. The limited use
 of collection after expulsion of the placenta is certainly due to the low
 yield of collection by gravity and to difficulty in handling the placenta,
 especially when the sterility of the collected blood must be guaranteed.
 Collection Systems
 Collection of placental blood from the umbilical cord in the medical field,
 both for diagnostic and therapeutic purposes and for research purposes, is
 currently performed with methods which can be traced back to two
 categories: "open-circuit" systems and "closed-circuit" systems. Both
 systems are used for collection both before and after expulsion of the
 placenta.
 a. "Open-circuit" Systems
 "Open-circuit" systems collect placental blood from the cut umbilical cord
 in containers which contain anticoagulant. This collection is performed by
 making the blood flow out directly into the collection container without
 connecting the end portion of the cord, which is placed loosely at the
 inlet of the container.
 Advantages
 The main advantage of this system is the absence of resistances to flow,
 since there are no bottlenecks or sudden decreases in cross-section of the
 cord and of its vessels.
 A second advantage is that the three vessels which are present in the
 umbilical cord are simultaneously pervious.
 Another advantage of the system is the possibility to express the umbilical
 cord during collection.
 Disadvantages
 Various scientific papers have demonstrated that "open-circuit" systems do
 not ensure sterility and are in fact associated with a particularly high
 incidence of microbial contaminations. The cut end portion of the
 umbilical cord in fact has a loose consistency, and since it is simply
 rested on the inlet of the collecting container it can easily escape from
 it, consequently contaminating said end portion in addition to losing
 blood, which contaminates the outside environment.
 Furthermore, the system inherently (being an "open" system) collects both
 the blood that flows out of the vessels of the cord and potentially
 contaminated liquids which flow along the outer surface of the umbilical
 cord; in particular, the maternal blood which is abundantly present along
 the outer surface of the placenta flows along the cord and mixes with the
 blood of the umbilical cord.
 This collection system furthermore exposes the blood to the air of the
 outside environment, thus facilitating contaminations by microorganisms
 which are present in the outside environment.
 Another drawback is the constant need for an operator assigned to keeping
 the umbilical cord in the correct position, particularly if repeated
 maneuvers for expressing and cleaning the cord are performed.
 b. "Closed-circuit" Systems
 Collection is performed by venipuncture of the cut and clamped end of the
 umbilical cord, after accurately disinfecting the entry point of the
 needle. The blood is collected in one or more syringes or in donation
 pouches.
 Advantages
 With respect to "open-circuit" systems, "closed-circuit" systems reduce the
 risks of microbial contamination of the collected blood and do not require
 continuous cleaning and disinfection of the cord.
 Another advantage is reduction of the contamination of the sample with
 maternal blood or other liquids which flow along the outer surface of the
 cord.
 Disadvantages
 "Closed-circuit" systems entail handling needles and therefore expose the
 operators to the risk of accidental punctures.
 Furthermore, the needle inserted in the umbilical vein is not stably fixed
 to it and can easily come loose, also in view of the highly dynamic nature
 of childbirth; this problem is particularly felt if collection is
 continued even after expulsion of the placenta. Accordingly, the
 continuous presence of an operator is required to ensure the correct
 placement of the needle during the various steps of collection.
 If the needle leaves its seat, blood contaminations and accidental
 punctures of the operator are possible, and repetition of venipuncture is
 furthermore required in order to continue collection.
 Owing to the limited cross-section of two of the three vessels of the
 umbilical cord (the arteries), collection is feasible only on one vessel
 (the vein), consequently reducing the collection potential.
 The flow of blood is also hindered by the sudden decrease in cross-section
 (the needle, no matter how large, can never have the same cross-section as
 the umbilical vein).
 The needle is inserted in an intermediate segment of the umbilical cord,
 upstream of the closure region; stasis of the blood downstream of the
 insertion point, with a consequent tendency to clotting, is thus
 inevitable; moreover, the blood which remains downstream of the insertion
 point is not recovered.
 SUMMARY OF THE INVENTION
 The aim of the present invention is to provide an improved method over the
 prior art cited above, for collecting blood or other fluids from the
 placenta or other organs or tissues for medical, scientific or other
 purposes.
 An object of the invention is to ensure maximum sterility of the sample by
 applying a procedure and using a closed-circuit apparatus which minimizes
 contact of the blood or fluid with potentially contaminated external
 factors or with the air.
 A further object of the present invention is to make the procedure
 accessible even to personnel who are not specifically trained. The
 procedure furthermore allows to limit the risks of human error on the part
 of the operator and, by using the suitable apparatus, to minimize the risk
 of injury to operators, since no exposed sharp or pointed objects are
 used.
 The above aims and objects, and other aims that will become apparent to
 those skilled in the art are achieved by a method for collecting blood
 from the placenta or another organ or tissue through the umbilical cord or
 a main vessel by means of a closed system which sequentially includes:
 1. Cleaning and disinfection of the portion of umbilical cord or vessel
 which is involved in the collection. The cleaning and disinfection system
 can be conveniently coupled to the system for connecting the umbilical
 cord or vessel.
 2. Cutting of the cord in a position which is suitable for subsequent
 coupling to the connection system. The cutting system can be conveniently
 coupled to the connection system of the umbilical cord.
 3. Connection of the umbilical cord or vessel by means of a connection
 system, optionally of the closed type, which can in turn be connected to a
 collection container; the connection system optionally includes a system
 for mixing the blood or other fluid with anticoagulant or other fluid or
 substance immediately after it has flowed out of the cord or vessel.
 4. Optional insertion of the placenta or other organ or tissue in
 containment means.
 5. Application of positive pressure to the placenta. The pressure can be
 uniform or, as an alternative, uneven, rhythmic, intermittent,
 centripetal, displacing or of one or more kinds, combined in various
 manners in order to mobilize the fluids from the more peripheral regions
 toward the main efferent vessels.
 6. Application of continuous or intermittent negative pressure, as
 required, to the connection or to the collection container in order to
 facilitate the outflow of blood or other fluids or to free the vessel from
 clots or other obstacles.

DESCRIPTION OF THE PREFERRED EMBODIMENTS
 In the procedure according to the invention, after childbirth 9, the
 umbilical cord is closed (clamped) 10 in two points and then cut 11 in a
 median position between the two closure points. The portion of the
 umbilical cord that is connected to the placenta protrudes freely from the
 mother's vagina 12.
 At this point, the end portion must be coupled to the collection container.
 The end portion of the umbilical cord is disinfected appropriately 13,
 optionally with a specifically designed disinfection system, is cut to
 size, if necessary, in order to adapt to the connection system 14, and is
 then coupled to the connection system 15. Said connection system (FIG. 2)
 is generally provided with suitable means for locking the cord, which
 prevents the cord from exiting from the system. The connection system is
 also provided with hermetic sealing means which isolate the cord portion
 from the outside environment at the inlet of said connection system,
 including any liquids which are present or flow on the outside wall of the
 umbilical cord. The sealing means are shaped appropriately and are
 constructed so as to not compress or choke the umbilical cord and in any
 case so as to avoid hindering in any way the collection of the blood or
 fluid. The connection system is then connected to the collection
 container, or the collection container can be an integral part of the
 connection system.
 The connection system 39 which comprises means 41 for forming a seal on the
 umbilical cord which delimit a portion of the umbilical cord toward the
 placenta 38a and an open end portion 38b, means 40 for locking the cord,
 elements for connection to a collection container 49 so as to delimit a
 hermetic closed space 50 which comprises an optional access path 48 for
 applying pressure or for introducing anticoagulant fluid or another fluid
 or for drawing part of the blood.
 The connection system, in one of its possible variations (FIG. 3), can also
 comprise in a single system, in addition to the vessel locking elements
 and to the sealing elements described above, cleaning and disinfection
 elements and cutting elements which are such that the operator, holding
 the end of the free portion of umbilical cord 12 with one hand, can
 disinfect the involved cord region, cut the cord to size and apply the
 connection system, which comprises both the locking elements and the
 sealing systems with a single operation 16.
 The connection system of FIG. 3 comprises means for disinfecting the end
 portion of the umbilical cord 43 and for cutting it 44.
 An additional variation of the connection system 17, illustrated in a
 possible non-limitative embodiment in (FIGS. 4A, 4B, 4C, and 4D), can
 comprise the various elements required for disinfecting, cutting and
 locking the cord and hermetic sealing means designed to be applied before
 separating the neonate from the mother. In this case, the operator can use
 the same instrument to close, disinfect, cut and lock the cord and place
 the sealing elements 17 directly after the birth of the neonate.
 The connection system of FIG. 4 comprises systems for closing or clamping
 45 the umbilical cord upstream 45a and downstream 45b of the cutting point
 of said cord; the clamping elements downstream of the cut or toward the
 neonate 45b are temporarily connected to the system and, after cutting the
 cord, can be disconnected to leave room for the collection container 42.
 Blood collection begins after applying the connection system. Before
 expulsion of the placenta, placental blood flows out owing to the pressure
 applied by uterine contractions to the placenta 18. After a variable
 period which lasts a few minutes, expulsion of the placenta occurs 19.
 During this step, the operator does not have to worry about the connection
 between the cord and the collection container, since the connection system
 has suitable locking means which ensure the stable connection between the
 cord, the connection system and optionally the collection container even
 during "dynamic" maneuvers such as expulsion of the placenta and transfer
 thereof to another location.
 After expulsion, the placenta is enclosed in a soft container (FIG. 5),
 optionally provided with an absorbent inner surface and an impermeable
 outer surface, which allows to avoid contaminating the outside environment
 and the operator 20 and is then subjected to pressure of various kinds by
 virtue of a suitable apparatus 21. During this second part of the
 collection 22, the flow of blood is ensured by the pressure applied
 appropriately by the specifically provided apparatus to the placenta. Said
 apparatus (FIG. 6) can be generally composed of elements which are
 suitable to apply pressures of various kinds to said placenta in suitable
 directions.
 Another possibility for improving the yield of placental blood collection
 is the optional application of negative pressure to the point where the
 blood flows out of the umbilical cord, so as to generate a suction effect
 23. The intensity of the suction is sufficient to facilitate the emptying
 of the cord and drainage of any obstacles, without however causing
 collapse of the vessels contained in the cord. The connection system in
 fact allows (FIG. 2, 3 and 4), by virtue of the hermetic sealing and
 pressure-tight systems, both on the umbilical cord and with the collection
 container, to create an enclosed space which can be pressurized. The
 negative pressure can be applied to the connection system both during the
 collection step before expulsion of the placenta 24 and during the second
 step after expulsion 25.
 The procedure described so far then entails, after expulsion of the
 neonate, the cutting, cleaning, disinfection and connection of the cord
 with a suitable connection system which comprises suitable means for
 locking and isolating from outside the open end portion of the cord. The
 connection system can be, or already is, connected to a collection
 container with suitable sealing means. This is followed by a first
 collection of blood from the cord prior to expulsion of the placenta.
 During expulsion of the placenta, the cord remains coupled to the
 connection system and to the collection container, since the cord is
 firmly fixed and isolated from outside by virtue of suitable locking and
 sealing means, respectively. After expulsion, the placenta is inserted in
 a suitable soft container which has suitable hermetic closure means; said
 container can be an integral part of the connection system by being for
 example rolled up and packed on the outer edges of the connection system
 and activated when required by unrolling along the umbilical cord and then
 around the placenta to be finally closed at the apex of said placenta. At
 this point, the placenta contained in the container is subjected to a
 suitable pressure, such as to facilitate the flow of blood toward the
 outside through the umbilical cord in the enclosed space. Where necessary,
 in order to optionally further improve collection or eliminate obstacles
 or clots of the umbilical cord, it is possible to apply negative pressure
 to the open end portion of the umbilical cord by virtue of an access path
 provided in the connection system or in the collection container. By
 virtue of the sealing elements which are present in the connection system
 and isolate the enclosed space which accommodates the open end portion of
 the umbilical cord from the space that contains the placenta, it is also
 possible to apply the pressure directly in the sealed container of the
 placenta.
 The described procedure allows, by means of a series of simple and safe
 operations, to maximize yield by virtue of sequential collection before
 and after expulsion of the placenta. The procedure also provides for the
 connection of the cord to a system which allows to keep all three vessels
 of the umbilical cord pervious, allowing them to contribute to the maximum
 yield of the collection. The operation provides for containment of the
 placenta and the cord from the initial steps, without using exposed
 pointed or sharp objects, ensuring the best possible safety of the
 operator. In the procedure, all the steps of collection are performed in a
 closed system, in which the blood does not make contact with the outside,
 thus preventing contamination of the sample. The procedure is easy and
 practical to use even for personnel who have not been trained
 specifically.
 The procedure is very quick, can be partially automated, is designed to
 minimize the burden of the operator and can be applied in dynamic working
 conditions, such as operating rooms, delivery rooms etcetera.
 In the system, as soon as the umbilical cord is cut, the placental portion
 of the umbilical cord from which the blood flows out is connected, after
 suitable disinfection, to the collection container by means of a closed
 connection system; said connection system eliminates the possibility of
 contaminating the blood, since it delimits a sterile closed region in
 which the blood passes from the umbilical cord to the collection
 container. The connection system, together with the container in which the
 placenta is placed, furthermore eliminate contaminations of the outside
 environment with placental blood, making the maneuver safe for the
 operator as well.
 The system also allows to optimize the collection volume, since it combines
 collection from the vein and from the two arteries with the safety and
 practicality characteristics typical of a closed system. The combined use
 of the apparatus for applying positive pressure furthermore allows, for
 the first time, to recover volumes of fluid which are up to 100% higher
 than those obtained with currently used procedures.