Note: Descriptions are shown in the official language in which they were submitted.
13~4189
CENTRIFUGAL FLUID PROCESSING SYSTEM AND METHOD
Field ~f the Invention
5The invention generally relates to systems
and methods for separating fluids by centrifugation.
More particularly, the invention relates to the cen-
trifugation of large volumes of fluids at relatively
high flow rates. In this respect, the invention also
10relates to systems and methods particularly well
- suited for the processing of cultured cells and super-
natant, such as in the fields of biotechnology and
adoptive immunotherapy.
Backaround of the Invention
15Many fluid processing techniaues entail the
centrifugation of large volumes of fluids. To mini-
mize processing times, these techniques often require
the use of relatively high flow rates. Increasingly,
such techniques are being used in the medical field.
20For example, in the areas of biotechnology
and adoptive immunotherapy, it is necessary to process
relatively large volumes of cultured cellular products
by centrifugation. Through centrifugation, cultured
cells are separated from the supernatant for the pur-
25pose of replacing/exchanging the culture medium; or
1334189
for providing a cell-free supernatant for the subse-
quent collection of antibodies or for subsequent use
as an additive to culture medium; or for the collec-
tion of concentrated cellular product.
In the area of adoptive immunotherapy, it
has been possible to process between 10 to 50 liters
of cultured LAK (Limphokine Activated Killer) cells at
a rate of 175 ml/min using conventional centrifugation
techniques and devices previously used in whole blood
processing. However, in the processing of TIL (Tumor
Infiltrating Lymphocytes), the volume of cultured
cells that must be processed is increased by an order
of magnitude to approximately 100 to 400 liters. Con-
ventional blood processing techniques and devices can-
not effectively deal with these large fluid volumes
and the attendant need to increase the processing
rates.
Purthermore, the necessarily high inlet flow
rates can lead to confused, turbulent flow conditions
within the centrifugation chamber. These flow condi-
tions are not desir~able, because they can interfere
with sedimentation and separation within the centrif-
ugal force field. Thus, despite the high inlet flow
rates, the overall effectiveness and efficiency of the
process suffers.
High inlet flow rates and resulting con-
fused, turbulent flow conditions can also result in a
non-uniform distribution of the fluid within the cen-
trifugation chamber.
Often, then, it is necessary to reduce the
inlet flow rate below the desired amount in the inter-
est of obtaining the flow conditions within the pro-
cessing chamber conducive to optimal separation.
Summarv of the Invention
The invention provides systems and methods
1334189
for centrifugally processing large volumes of fluid at
relatively high flow rates without sacrificing separa-
tion efficiencies or damaging the end product.
In one aspect, the invention provides a cen-
trifugal processing system and method in which a cen-
trifugal force field is developed within a chamber.
As the fluid to be processed is introduced into the
chamber, it is directed away from the region of the
chamber where the largest centrifugal (or "G") forces
exist. The fluid is also preferably conveyed into the
force field in a generally uniform stream. As used
herein, the term "generally uniform" describes a flow
condition in which turbulence is reduced or eliminated
to the fullest extent possible.
In accordance with this aspect of the inven-
tion, the system and method establish, upon the entry
of high velocity fluid into the centrifugal field,
generally uniform flow conditions conducive to effec-
tive separation. The system and method also direct
the fluid in a way that maximizes the effective sur-
- face area of the centrifugation chamber for separa-
tion. Effective separation can thereby be achieved at
high inlet flow rates.
Preferably, the system and method embodying
the features of the invention also create within the
chamber a region where the higher density materials
collect, while allowing the supernatant to freely flow
out of the chamber.
In another aspect of the invention, the cen-
trifugation chamber takes the form of a tube or
envelope. In this embodiment, a passage is formed
within the tube ad~acent to its inlet end. All fluid
entering the tube is directed through this passage and
into the centrifugal force field. The passage creates
a generally uniform stream of fluid having reduced
4 1334189
turbulence or being essentially free of turbulence.
This stream is directed and dispensed uniformly into the
region of the tube where the least centrifugal forces
exist.
Various aspects of the invention are as follows:
A centrifugal chamber for positioning within a
rotating field and for centrifugally processing a fluid
suspension into component parts, the chamber comprising:
oppositely spaced first and second exterior walls
defining a chamber having an interior processing region
with an inlet, the first exterior wall, when positioned
within the rotating field, being disposed closer to the
rotational axis than the second exterior wall to define
within the interior processing region a low-g area
adjacent the first exterior wall and a high-g area
adjacent the second exterior wall,
inlet conduit means of a given cross sectional area
for conducting the fluid suspension to be processed to
the chamber, and
wall means forming a fluid receiving area within
the inlet of the interior processing region in
communication with the inlet conduit means, the fluid
receiving area including an interior wall that isolates
the fluid receiving area from the interior processing
region except for an exit passage that has a cross
sectional are greater than the cross sectional area of
the inlet conduit means and that extends transversely
across the exterior wall for dispensing the fluid
suspension conducted by the inlet conduit means only
into the low-g area of the processing region in a
generally uniform flow free or essentially free of
turbulence.
A centrifugal processing system comprising
a rotor rotatable about an axis, and
a processing chamber carried by the rotor in an
arcuate path about the rotational axis, the processing
chamber having oppositely spaced first and second
4a l 3341 8~
exterior walls that enclose an interior processing
region with an inlet at one arcuate location and an
outlet at another accurately spaced location on the
rotor, the first exterior wall, when positioned within
the rotating field, being disposed closer to the
rotational axis than the second exterior wall to define
within the interior processing region a low-g area
adjacent the first exterior wall and a high-g area
adjacent the second exterior wall,
inlet conduit means of a given cross sectional area
for conducting the fluid suspension to be processed to
the processing chamber, and
wall means forming a fluid receiving area within
the inlet of the processing region in communication with
the inlet conduit means, the fluid receiving area
including an interior wall that isolates the fluid
receiving ~rea from the interior processing region
except for an exit passage that has a cross sectional
area greater than the cross sectional area of the inlet
conduit means and that extends transversely across the
first exterior wall for dispensing the fluid suspension
conducted by the inlet conduit means only into the low-g
area of the processing chamber in a generally uniform
flow free or essentially free of turbulence.
A centrifugal processing method for separating the
higher density components of a fluid from the lower
density components of the fluid comprising the steps of:
providing a chamber having an interior processing
region and an inlet,
developing a centrifugal force field within the
- interior processing region to therein create a low-g
area and a high-g area,
conveying the fluid to be processed through an
inlet path of a given cross sectional area into a
receiving area within the inlet to the processing
region, the receiving area having an interior wall that
isolates the interior processing region from the flow
4b 13341 8q
conditions created by conveying the fluid into the
receiving area,
further conveying the fluid from the receiving area
through an exit passage that has a cross sectional area
greater than the cross sectional area of the inlet path
and that dispenses the fluid in a generally uniform flow
free or essentially free of turbulence only into the
low-g area of the processing chamber.
Other features and advantages of the lnven-
tion will become apparent upon considering the accom-
panying drawings, description, and claims.
Brief DescriDtion of the Drawinas
Fig. 1 is a schematic side view, fragmented
and partially in section, of a centrifugal processing
system embodying the features of the invention;
Fig. 2 is a top view of the centrifugal pro-
cessing system taken generally along line 2-2 in Fig.
l;
Fig. 3 is an enlarged fragmented top view of
the processing tube or envelope of the fluid proces-
sing set associated with the system shown in Fig. l;
Fig. 4 is a side view of the processing tube
or envelope taken generally along line 4-4 in Fig. 3;
Fig. 5 is an exploded perspective view of
the processing tube shown in Fig. 3 showing the asso-
ciated flow control means;
Fig. 6 is an enlarged schematic view, frag-
mented and broken away in section, of the processing
tube or envelope shown in Figs. 3 to S illustrating
the flow of fluid through the tube or envelope when it
- is in use in a centrlfugal fleld;
~ - Fig. 7 is a greatly enlarged schematic view,
fragmented and ln section, of the collection of higher
density materials in the tube or envelope shown ln
Flg. 6;
Flg. 8 ls a centrifugal fluid processlng
system embodylng the features of the invention and
lntended to be use ln the harvestlng of cell cultures
on a large volume basls; and
1 3341 8q
Fig. 9 is an alternate embodiment of a cen-
trifugal fluid processing system embodying the fea-
tures of the invention.
Descri~tion of the Preferred Embodiments
S A centrifugal fluid processing system 10 em-
bodying the features of the lnvention is shown in Fig.
1. The system 10 includes a centrifuge 12 and an
associated fluid processing æet 14. In the illus-
trated and preferred embodiment, the set 14 is dispos-
able, intended to be used once and then discarded.
The system 10 can be used to process many
different types of fluid. As will become apparent,
the system 10 is capable of efficiently processing
large volumes of fluid at relatively high flow rates.
At the same time, the system 10 is well adapted to
handle special fluids containing living cells or deli-
cate organisms, such as blood or cultured cell suspen-
sions, both on a clinical basis and an industrial
basis. For this reason, the system 10 is particularly
well suited for use in the medical field. For this
reason, the system 10 will be described as being used
in thls particular environment.
The centrifuge 12 can be variously con-
structed. However, in the illustrated embodiment, the
centrifuge 12 is shown to incorporate the principles
of operation disclosed in Adams U. S. Patent No. Re
29,738.
In this arrangement (as best shown in Fig.
1), the centrifuge 12 includes a processing assembly
16 and a rotor assembly 18 each of which independently
rotates about the same axis 20. The processing assem-
bly 16 is connected to a first drive shaft 22. The
rotor assembly 18 is connected to a second drive shaft
28. The second drive shaft is driven via a suitable
pulley assembly 24 by a drive motor 26. The first
1 S34 1 8~
drive shaft 22 is driven by a suitable pulley assembly
30 associated with the second drive shaft 28.
The pulley assemblies 24 and 30 are conven-
tionally arranged to cause the processing assembly 16
S to rotate in the same direction as and at twice the
rotational speed of the rotor assembly 18. Examples
of this type of construction are more fully disclosed
in Lolachi U. S. Patent 4,113,173.
As can be best seen in Figs. 1 and 2, the
processing assembly 16 includes an inner processing
area 32. The processing area 32 takes the form of an
arcuate slot or channel. The slot 32 can be config-
ured in various ways, depending upon the intended use
of the system. In the illustrated embodiment (best
shown in Fig. 2), the slot 32 is generally equally
radially spaced about the rotational axis 20 shared by
processing assembly 16 and rotor assembly 18.
With further reference now to Figs. 3 to 5,
the fluid processing set 14 includes an envelope or
tube 34 defining a hollow interior chamber 36 having
an inlet end 38 and an outlet end 40. The tube 34 is
intended to be inserted into the processing slot 32
(see Figs. 1 and 2). As will be soon described in
greater detail below, the intended centrifugal separa-
tion of the processed fluid occurs within the interior
chamber 36 of the tube 34 due to centrifugal forces
created during rotation of the processing assembly 16.
The tube 34 is can be made from either a
flexible or rigid material. When flexible, the tube
34 can be readily fitted into the slot 32 to there
conform to the particular configuration of the slot
32. When rigid, the tube 34 would be preformed to
match the particular configuration of the slot 32. In
the illustrated embodiment, which contemplates use of
the system 10 in the medical field, the tube 34 is
1334189
made from a flexible medical grade plastic material,
such a polyvinyl chloride.
As best shown in Fig. 1, the fluid proces-
sing set 14 further includes inlet tubing 42 for con-
veying fluid into the inlet end 38 of the tube chamber
36 for centrifugal separation. Likewise, the set 14
includes outlet tubing 44 for conveying fluid constit-
uents from the outlet end 40 of the tube chamber 36
after processing.
In the illustrated embodiment, there are two
inlet tubes 42 and three outlet tubes 44 (see Fig. 3).
Of course, the number of tubes can vary according to
the intended use and function of the system 10.
In the illustrated embodiment, the inlet and
outlet tubing 42 and 44 are made from flexible medical
grade plastic material and are joined to form a mul-
tiple lumen umbilicus 46. As best shown in Fig. 1,
the umbilicus 46 is suspended from a point above and
axially aligned with the rotational axis 20 of the
centrifuge 12 by means of a clamp 48 attached to a
support arm 50. From this point, the umbilicus 46
extends generally downwardly and radially outwardly,
passing against a guide arm 52 carried by the rotor
assembly 18. From there, the umbilicus 46 extends
generally downwardly and radially inwardly and then
upwardly through the hollow center of the drive shaft
22 into the processing assembly 16.
This looping arrangement of the umbilicus
46, coupled with the differing rotational rates of the
processing assembly 16 and the rotor assembly 18 as
just described, prevents the umbilicus 46 from becom-
lng twisted during operation of the centrifuge 12.
The use of rotating seals between the flxed and rotat-
ing parts of the system 10 is thereby avoided. How-
ever, it should be appreciated that the invention is
133418q
applicable for use in other types of centrifugal sys-
tems, including those employing rotating seals.
Once the tube 34 is located in the proces-
sing area 32 and filled with fluid, the rotation of
the processing assembly 16 will create a centrifugal
force field F (see Fig. 2) effecting the contents of
the tube chamber 36. This force field F will create
a "High G Region" 54 and a "Low G Region" 56 within
the chamber 36. As shown in Fig. 2, the "High G Re-
gion 54" is located adjacent to the outer wall of the
chamber 36, where the force field is farthest away
from the rotational axis and the contents of the cham-
ber 36 are subjected to the highest rotational (or
"G") forces. The "Low G Region 56" is located adja-
cent to the inner wall of the chamber 36, where the
force field is nearer to the rotational axis and the
contents of the chamber are subjected to lesser rota-
tional (or "G") forces. As best shown in Figs. 6 and
7, higher density materials present in the processed
fluid (designated 101 in Figs. 6 and 7) will migrate
under the influence of the force field F toward the
High G Region 54, leaving the less dense materials and
supernatant (designated 115 in Figs. 6 and 7) behind
in the Low G Region 56.
To obtained the desired flow rate condi-
tions, the fluid to be processed is introduced into
the tube chamber 36 using a suitable in line pumping
mechanism 58. In the illustrated embodiment (see Fig.
1), the pumping mechanism takes the form of a peris-
taltic pump 58 situated upstream of the tube chamber
36.
In Fig. 8, the set 14 as just described is
shown particularly configured for use to harvest TIL
cells. In this procedure, cultured TIL cell solution
filling approximately 70 to 260 three liter bags 60,
1334189
each filled with about 1-1/2 liters of æolution, is
centrifugally processed to remove the æupernatant and
obtain concentrated TIL cells (which presently con-
sists of approximately 2 x lOt1 cells occupying a vol-
ume which ranges between 220 to 400 ml).
In this arrangement, 5-lead and 10-lead man-
ifold sets 62 are used to interconnect the many supply
bags 60 to a single inlet line 64. The cultured cell
fluid is then conveyed into a reservoir bag 66, using
the supply pump 68, and then conducted into the tube
34, using the processing pump 58.
In this arrangement, the tube 34 is approxi-
mately 32 inches long and 3 inches wide. The interior
surface area of the tube 34 is approximately 200
square inches.
During centrifugation, the TIL cells are
separated from the culture medium (which constitutes
the supernatant). The supernatant is collected in
large volume containers 72. Afterwards, the concen-
trated TIL cells are transferred to a collection con-
tainer 74 for administration to the patient.
In this and other applications, where rela-
tively large volumes of fluid are to be processed, it
is desirable to maximize the inlet flow rate of the
fluid, as this will shorten the overall processing
time. In the case of a TIL procedure, a nominal proc-
essing rate of at least 1.5 liters per minute is
attained. However, with the system 10 illustrated, it
is believed that the processing rate can be increased
upwards to about 4 liters per minute. This rate is
significantly higher than the nominal processing rates
conventionally used for blood processing (about 50
ml/min) or conventionally used for TIL cell harvesting
(about 175 ml/min).
Use of these relatively high inlet flow
-
lo - 1 3341 89
rates can pose processing problems. In particular,
such high rates can lead to confused, turbulent flow
conditions within the tube chamber 36. These turbulent
or otherwise confused, non-unlform flow conditions can
interfere with sedimentation and separation within the
centrifugal force field F, lowering the overall effec-
tiveness and efficiency of the process.
High inlet flow rates and attendant con-
fused, turbulent flow conditions can also result in a
non-uniform distribution of the fluid within the tube
chamber 36. To maximize the effective surface area
along which separation occurs, the incoming fluid
should preferably enter in the Low G Region 56 as soon
as possible after entering the tube 34. The fluid
lS components are thereby exposed to the full extent of
the centrifugal force field F for the longest period
of time. However, high inlet flow rates can spray or
disperse the incoming fluid lndiscrlmlnately lnto both
the High and Low G Regions 54 and 56 of the tube 34.
This, too, lowers the overall effectiveness and effi-
ciency of the process.
To optimize the effectiveness of separa-
tion at high inlet flow rates, the invention provides
a fluid processing system 10 that includes means 76
located adjacent the inlet end of the tube chamber 36
for directing incoming fluid away from the High G
Region 54 and toward the Low G Region 56 of the cham-
ber 36 in a generally uniform flow having reduced tur-
bulence or being generally free of turbulence. Pre-
ferably, the unlform flow constitutes a relatively
thin stream filling the entire effective surface area
of the Low G Reglon 56 adjacent to the inlet end of
the chamber 36.
In accordance with the invention, the means
76 therefore establishes, upon the entry of high velo-
1 334 1 89
city fluid into the centrifugal field F, the desired
flow conditions for effective æeparatlon. The means
76 also directs and dispenses the fluid in a manner
that maximizes the effective surface area of the tube
chamber 36 for separation. Due to the invention,
effective separation can be achieved, even at high in-
let flow rates.
The means 76 can be variously constructed.
One embodiment is shown-in Figs. 3 to 5. In this
arrangement, the means 76 is part of a port block
assembly 78 situated within the inlet end 38 of the
tube chamber 36. The assembly 78 lncludes top,
bottom, and side walls 80; 81; and 82 defining an open
interior 84. The assembly 78 also includes a first
end wall 86 closing the adjacent end of the interior
84. One or more inlet ports 88 are formed on this end
wall 86. The inlet tubing 42 is attached to these
ports 88 to introduce fluid into the open interior 84
of the assembly 78.
In this arrangement, the means 76 comprises
a partial second end wall 90 located on the end of the
port block assembly 78 opposite to the end wall 86 on
which the inlet ports 88 are situated. This partial
end wall 90 extends from the top wall 80 toward the
bottom wall 81, terminating a short distance therefrom
to there define a flow passage 92 communicating with
the open interior 84 of the assembly 78. As will be
described in greater detail below, fluid introduced
into the open interior 84 of the port block assembly
78 (via the inlet ports 88) is directed into the cen-
trifugal force field through the flow passage 92.
As best shown in Fig. 4, the port block
assembly 78 is situated within the inlet end of the
tube chamber 36 with the flow passage 92 extending
longitudinally across the entire interior surface of
- 12 -
1 334 1 89
the tube chamber 36 which, in use, becomes the Low G
Region 56.
To assure that the interior surface of the
tube 34 becomes the Low G Region 56 when situated
within the processing area 32, a guide key 94 is pro-
vided on the port block assembly 78 which mates with
a groove 96 in the processing area 32 (see Fig 2) when
the tube 34 is properly oriented.
The system lO further includes means 98
defining a region 100 for collecting high density ma-
terials within the tube chamber 36. In the embodiment
shown in Figs. 2 to 5, the means 98 includes a dam
assembly 102 situated within the tube chamber 36 down-
stream of the port block assembly 78. The dam assem-
bly 102 may be variously constructed. In the illus-
trated embodiment, the dam assembly 102 is part of
another port block assembly as previously described.
The assembly 102 includes top and bottom walls
103/104, side walls 105, and an end wall 106.
In this arrangement, the dam assembly 102
comprises a partial end wall 108, which like the means
76 associated with the port block assembly 78, forms
another flow passage 110 through which fluid must pass
to exit the tube chamber 36.
The length of the end wall 108 associated
with the dam assembly 102 can vary. It can be the
same as or different than the end wall 90 of the port
block assembly 78, depending upon the nature and type
of collection area or areas sought to be established
within the tube chamber 36. The sedementation of
higher density materials in the region 100 is also
effected by the fluid flow rate, the RPM of the cen-
trifuge, and the interior thickness of the tube cham-
ber 36. These variables can be adjusted to alter the
collection characteristics of the tube 34.
- 13 -
1 334 1 89
It should also be appreciated that multiple
dam assemblies of varying lengths and spacing can be
used to create multiple separation and sedimentation
zones within the tube chamber 36.
As shown in Figs. 6 and 7, and as will be
described in greater detail below, the higher density
materials (designated 101 in Figs. 6 and 7) migrating
toward the High G Region 54 of the chamber 36 will
collect within the area 100 bounded by the partial end
wall 90 of the port block assembly 78 and the partial
end wall 108 of the dam assembly 102.
In the embodiment shown in Figs. 3 to 5, the
dam assembly 102 is located in the outlet end 40 of
the tube chamber 36, and outlet ports 112 are accord-
ingly formed on the end wall 106, as in the port block
assembly 78. However, it should be appreciated that
the dam assembly 102 can be located within the tube
chamber 36 at a location upstream of the outlet end 40
of the chamber 36 (as shown in Fig. 6), in which case
the end wall 106 would be free of ports. In this
arrangement, a separate port block assembly (not
- shown), without a partial end wall, would be used at
the outlet end 40 of the tube chamber 36.
The port block assembly 78 and the dam
assembly 102 can be made of various materials. In the
illustrated embodiment, both are injection molded
plastic parts that are located and sealed within the
confines of the tube chamber 36 by heat sealing, sol-
vent sealing, or similar techniques.
The dimensions of the flow passages 92 and
110 can vary according to the type of fluid being pro-
cessed and the flow conditions desired. In the par-
ticular embodiment shown in Fig. 8, the flow passages
92 and 110 are each about 3 inches wide (the same
width as the associated tube) and about .025 inch in
-
lS~418q
height. The passages 92 and 110 therefore comprises
restricted flow passages.
Another embodiment of the means 76 for dir-
ecting incoming fluid toward the Low G Region 56 is
shown in Fig. 9. In this arrangement, the means 76
takes the form of a ridge 114 formed within the out-
side (High G) side of the processing area 32 of the
assembly 16. When the tube 34 is positioned within
the processing area 32 (as shown in Fig. 7), the ridge
114 presses against the exterior of the outside wall
of the tube 34, thereby forming a passage 92 like that
formed by the partial end wall 90 of the port block
assembly 78. Preferably, a recess 116 is formed in
the inside (Low G) side of the processing area 32 ra-
dially across from the ridge 114 to facilitate inser-
tion and removal of the tube 34 and to better define
the passage 92.
As also shown in Fig. 9, the means 98 for
defining the collection area 100 for higher density
materials can also take the form of a ridge 118 and
associated recess 120 formed along the walls of the
processing area 32 of the centrifuge 12.
A centrifugal processing method which embod-
ies the features of the invention is shown in Figs. 6
and 7. This process will result by the operation of
the above described port block assembly 78 and dam
assembly 102 when the tube chamber 36 is exposed to
the centrifugal field F. However, it should be appre-
ciated that the process can be achieved by other means
as well.
In this method, as the fluid to be processed
is introduced into the centrifugal force field F, it
is directed away from the region of the chamber 36
where the largest centrifugal (or "G") forces exist.
Furthermore, the fluid is directed and dispensed into
- 15 -
13341~
the force field as a generally uniform stream (desig-
nated by arrows and number 111 in Figs 6 and 7) having
reduced turbulence of being essentially free of turbu-
lence.
s Referring specifically now to Figs. 6 and 7,
incoming fluid entering the port block assembly 78
(via the ports 88) is immediately confined within the
open interior 84. Turbulent flow conditions occasioned
by the entry of fluid into the chamber 36 (indicated
by swirling arrows 113 in Figs 6 and 7) are thereby
effectively confined to this interior area 84 and iso-
lated from the remainder of the tube chamber 36.
The fluid confined within the interior area
84 is directed by the partial end wall 90 away from
the High G Region 54 and out into the tube chamber 36
via the passage 92. By virtue of the shape of the
passage 92, the fluid is directed and dispensed in a
generally uniform stream 111 extending across the Low
G Region 56 of the tube chamber 36.
Optimal conditions for sedimentation and
separation are thereby quickly established. As a
result, the higher density materials 101 migrate due
to the force field F toward the High G Region 54. The
remaining supernatant (designated by arrows and number
115 in Figs. 6 and 7) continues to flow uniformly
along the Low G Region 56 toward the outlet end 40 of
the tube chamber 36.
The process also creates within the chamber
36 a region 100 where the higher density materials 101
collect, while allowing the supernatant 115 to flow
freely out of the chamber 36. As can be best seen in
Fig. 6, the higher density materials 101 migrating
toward the High G Region 54 of the chamber 36 collect
within the area 100 bounded by the partial end wall 90
of the port block assembly 78 and the partial end wall
- 16 -
1 334 1 8q
108 of the dam assembly 102. At the same time, the
supernatant, which is free of the higher density ma-
terials 101, passes through the passage 110 of the dam
assembly 102 and exits the outlet end 40 of the tube
chamber 36.
EXAMPLE 1
A tube 34 embodying the features of the in-
vention was used in association with a set as gener-
ally shown in Fig. 8 and an Adams-type centrifuge to
harvest human red blood cells from a saline suspen-
sion. Three runs were conducted.
In the first run, the suspension had an
original red blood cell concentration of 1.27 x 107
per ml. This suspension was centrifugally processed
through the tube at a flow rate of 1800 ml/min at 1600
RPM. During processing, concentrated red blood cells
were collected at processing efficiency of 94.9%.
In the second run, the original suspension
concentration was 1.43 x 107 red blood cells per ml.
During centrifugal processing at a flow rate of 1000
ml/min at 1600 RPM, concentrated red blood cells were
collected at a processing efficiency of 95.7%.
In the third run, the original suspension
concentration was 1.33 x 107 red blood cells per ml.
During centrifugal processing at a flow rate of 1800
ml/min at 1600 RPM, concentrated red blood cells were
collected at a processing efficiency of 91.5%.
EXAMPLE 2
A tube 34 embodying the features of the in-
vention was used in association with a set as genera-
lly shown in Fig. 8 and an Adams-type centrifuge to
harvest TIL cells from suspension.
During the procedure, 24,559 ml of cultured
TIL cell suspension was processed through the tube a
flow rates varying between 500 to 1500 ml/min at 1600
-
- 17 -
lS3418q
RPM. 445 ml of concentrated TIL cells were obtained.
Approximately 564.9 x 10~ TIL cells were
contained in the suspension prior to processing. Dur-
ing processing, approximately 462.8 x 10~ TIL cells
were collected, for a processing efficiency of 82%.
TIL cell viability of 73% was measured prior
to processing. TIL cell viability of 73% was measured
after processing.
Lytic activity of the TIL cells prior to
processing was 5.4%. After processing, the lytic
activity was 4.3%, which does not constitute a
statistically significant difference.
The foregoing examples clearly illustrate
the ability of a processing system made and operated
in accordance with the invention to efficiently pro-
cess large volumes of cellular suspensions at rela-
tively high fluid flow rates. Example 2 further dem-
onstrates that processing occurs without biological
damage to the cellular components.
Various features of the invention are set
forth in the following claims.
