Reagents comprise No. According to the invention, cerebroprotein hydrolysate is added in the No. By using the stem cell sample density separating medium of the invention, the separation and recovery rate of target stem cells are improved by about percent, the human marrow, umbilical cord blood or peripheral blood samples can be effectively saved, and on the basis of maintaining high survival rate, the separation amount, the recovery rate and the use efficiency of stem cells are greatly improved.
The invention is an unprecedented breakthrough in clinical treatment applying stem cells. The isolated stem cell density of the sample solution according to claim I, wherein: the brain protein hydrolyzate is selected from one of the following products: Zhunzi H, Zhunzi H, H or RESEARCH Zhunzi quasi-word H; brain component content of the protein hydrolyzate is O.
Stem cell sample liquid density separation according to claim 6, wherein: said liquid No. A method of isolating stem cells according to claim 8, wherein: said bone marrow of step I of the, umbilical cord blood or peripheral blood samples, anticoagulated, diluted under sterile conditions required to complete its collection, anticoagulation are: bone marrow blood collection: with superior iliac spine, the method Guchuan local anesthesia, using the collection of bone marrow collection broach, the bone marrow collection needle used in the first extraction 5ml sodium citrate.
A method of isolating stem cells according to claim 9, wherein: 1 the step I , the harvested bone marrow after anticoagulated blood, umbilical cord blood or peripheral blood sample is pretreated with I liquid volume number ratio of the I: 1, to give a diluted sample; 2 in said step 2 , the sample No.
CN CNA en Human marrow, cord blood or peripheral blood stem cells treating kit and stem cells separating method. CN CNB en Stem cell sample density separating medium and stem cell separation method for human marrow, umbilical cord blood or peripheral blood. CN CNA en.
CN CNB en. CNA en. Application of human stem cell composition in preparing medicine for treating AIDS. Use of human stem cell composition in preparation of medicaments for treating alzheimer's disease. Use of human stem cell composition in preparing medicament for treating premature ovarian failure or uterine diseases.kirtihospital.com/a-million-windows.php
Cell Separation - 1st Edition
Method for separating, purifying and culturing human blood and marrow dendritic cells. Kits and methods for processing stem cells from bone marrow or umbilical cord blood. Kit for allogeneic peripheral blood mononuclear cell separation in vitro and application method of kit. More information about this seller Contact this seller 7. Condition: Near Fine. Dust Jacket Condition: Very Good-. First Printing Stated. More information about this seller Contact this seller 8. Condition: Used: Good. More information about this seller Contact this seller 9.
Published by Academic Press Inc From: Anybook Ltd. Lincoln, United Kingdom. About this Item: Academic Press Inc, Volume 1. This book has hardback covers. In good all round condition. No dust jacket. Please note the Image in this listing is a stock photo and may not match the covers of the actual item,grams, ISBN More information about this seller Contact this seller Condition: Fair. Volume 4. Ex-library, With usual stamps and markings, In fair condition, suitable as a study copy. Please note the Image in this listing is a stock photo and may not match the covers of the actual item,grams, ISBNX.
Condition: Poor. Volume 2. In poor condition, suitable as a reading copy. Published by Academic Press Inc, Gebundene Ausgabe. Condition: Gut. Barcode und Inventarisierungsnummer ; in der Regel foliiert Umschlag aus selbstklebender Folie. Tagesaktueller, sicherer und weltweiter Versand.
Published by Academic Pr, New York No Jacket. First Edition. The book is an ex-library book with the usual stamps etc. As the number of fields coupled with FFF is large the selection criteria are wide-ranging: size, shape, density, rigidity, subcellular ultrastructure, etc. As a result of low shear rates and minimal handling of cells FFF techniques typically have high recovery or yield. Recently, FFF has been directed at the separation of stem cells from clinical samples.
Vykoukal et al. For further reading on field-flow fractionation, an extensive review of recent trends was written by Roda et al. Microstructure protrusions planar and lateral to flow have been used for rapid mixing in lab-on-a-chip applications. Flow past static structures creates lateral flow components and flow alteration within the channel that induce mixing [ 59 , 60 ]. However, they have also been proven useful for entraining cells and performing size-, deformability-, and density-based cell separation.
Several mechanisms for particle separation in these helical flows have been proposed. Geometries have been designed for binary separation of particles of different sizes and for concentrating or focusing all particulate matter to a single stream. Likely, particles are entrained in helical flows and are acted on, differentially, by gravity and sterics size differences restrict movement around microstructures. These microstructures are capable of precise control of cell distribution within a microchannel enabling the resolution of cells with diameter differences as small as 7.
That is, size differences result in deterministic placement into fluid streamlines in an analysis or collection region. However, these cells were not collected off-chip. Recently, Choi et al. Herringbone grooves create a flow pattern suitable for segregating similarly sized particles based on density. A density resolution of 0. While focusing for massively parallel imaging was demonstrated, outlet configurations for actually separating cells or particles in this device were not. We recently reviewed inertial microfluidics and its applications to label-free sorting and direct the reader there for a more complete background [ 63 ].
Briefly, in the upper range of flow rates explored in microfluidic systems Reynolds numbers in the range of 1— are common. In this range inertial effects become significant, and the assumption that particles follow fluid streamlines leads to incorrect results. For a circular pipe, initially randomly dispersed, suspended particles concentrate to a narrow band at approximately 0.
This focusing phenomenon has been described as a balance of two inertial lift forces: 1 the shear gradient lift and 2 the wall effect lift. Furthermore, the number and location of equilibrium positions may be manipulated by controlling channel geometry [ 65 , 66 ]. A second inertial effect in microfluidic systems arises in curving channels such as arcs and spirals.
Higher momentum fluid at the center of a channel displaces lower momentum fluid near channel walls as it is driven around a curve [ 67 ]. This results in counter-rotating vortices perpendicular to primary channel flow which may also influence particle positions [ 68 ]. Cells can be entrained in this secondary flow and may be dragged perpendicular to the primary flow. As in other techniques there can be equilibrium and kinetic inertial separations. For efficient separation based on differential equilibrium positions two conditions must be met: 1 both the selected and unselected particles must be accurately focused, and 2 their equilibrium positions must be different.
Inertial lift forces depend on particle size, but equilibrium positions in straight channels are roughly the same for all particle sizes so long as the length of the channel gives sufficient time for particles to migrate to these equilibriums. Creation of distinct equilibrium positions requires an additional, size-dependent force on the order of the inertial lift force.
The Dean drag force found in spiral or asymmetrically curving channel geometries is such a force. Equilibrium position-based separation was explored for membrane-free filtration, enriching particulate matter in suspension. In these devices suspended particles are driven through a microchannel by a syringe pump. Initially randomly distributed throughout the channel, particles above a critical size migrate to equilibrium positions. Strategically placed outlet channels or bifurcations separate fluid with concentrated particles from fluid with few to no particles [ 69 ].
Seo et al. This type of enrichment has been replicated in a number of microchannel designs. Kuntaegowdanahalli et al. Kinetic, inertial separation is possible by utilizing differences in the lateral lift forces as they depend on particle size. Wu et al. Blood cells felt a larger lift force than bacteria, resulting in significant differences in position within the channel cross section.
Inertial microfluidic separation typically involves a dilution, either prior to injection or within the device, as interparticle interactions lead to defocusing. Furthermore, it is anticipated that increased viscosity, due to large cellular content, would increase the channel length required to achieve focusing.
However, this restriction is compensated for by high volumetric flow rates, surpassing most other microfluidic technologies by orders of magnitude. As such, inertial microfluidic separations should be considered for applications which would benefit from the throughput and is being commercially developed for water filtration Palo Alto Research Center.
Lastly, their independence from external force fields eases their integration into massively parallel systems [ 65 , 73 ]. Huh et al.
Technologies for circulating tumor cell separation from whole blood
Unlike the inertial separation described in the previous section, gravitational and sedimentation methods depend on the density of the particle, rather than the density of the fluid. A randomly distributed particle suspension is injected into the device parallel to gravity, and particles are hydrodynamically focused in one dimension by sheath flows. Separation of particles with different radii is amplified by hydrodynamic effects: the channel gradually widens asymmetrically such that flow near the bottom of the channel is angled downward, assisting migration of larger radius particles.
Microdevices that mimic the microvasculature. Copyright American Chemical Society. Cell separation from plasma is another common objective.
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The plasma skimming phenomenon has been exploited in vitro for separation of cell-free plasma. Faivre et al. They demonstrated the cell-free layer could be enhanced by a sudden channel expansion. Sollier et al. Important parameters include the suspension viscosity adjusted by dilution , the shape and size of the expansion, and flow rate.
At these high flow rates inertial effects may affect the plasma skimming. Bifurcation law was also manipulated to achieve highly efficient plasma separation across a wide range of hematocrits. Successive branches off the main channel remove plasma Fig. Biomimetic separation, comprised of a number of disparate techniques, may be applied in a number of ways.
The operational flow rates of the plasma-skimming devices tested by Faivre et al. The devices produced by Shevkoplyas et al. Magnetic activated cell sorting is a label-based, continuous, passive separation method. However, there are microfluidic systems which employ the intrinsic magnetophoretic properties of hemoglobin, found in eryrthrocytes, for separation.
Unpaired electrons in deoxygenated and methemoglobin make them paramagnetic while these electrons form covalent bonds in oxyhemoglobin making it diamagnetic. Zborowski et al. Han and Frazier created several devices for magnetophoretic separation [ 80 , 81 , 82 ]. However, the separation force in these systems decayed with distance into flow. Furlani provided a theoretical model of a continuous flow, gravity-driven microfluidic systems for separation of WBCs and deoxygenated RBCs which instead uses an array of soft-magnetic elements which are magnetized by a bias field [ 83 ].
Therefore, they move in opposite directions perpendicular to primary flow. The proposed device has not been experimentally validated. Inherent in throughput calculations for these devices is a dilution step to chemically prepare deoxygenated hemoglobin red blood cells. Separate outlets contained Huang et al. The first module, as described above, was a DLD microfluidic device for depleting the smaller, non-nucleated red blood cells. Subsequently, a magnetic column, with a magnetic field of 1. As with other techniques, separation efficiency, purity, and throughput were at odds.
The commercial development Artemis Health of this technique and its unique application for noninvasive diagnostics suggests promise. Aqueous two-phase systems ATPS are a technique used for separation of biological materials. Briefly, separation is based on the differential affinity of biological materials for either of two immiscible polymers usually polyethylene glycol PEG or dextran or their interface.
Separation criteria include surface properties and net charge. Salts such as lithium sulfate may be added to change affinity or maintain physiological osmolarity. However, ATPS cell separation in the macroscale has many drawbacks. Yamada et al. Microfluidic ATPS can also be used to separate live and dead cells [ 86 ]. SooHoo and Walker [ 20 ] developed a device for leukocyte enrichment from whole blood by manipulating the differential affinity of white and red blood cells for PEG or dextran phases or their interface.
They achieved a 9. The mechanism for these specific interactions has not been thoroughly evaluated. Dilution is limiting and syringe pump fluctuations affecting interface stability or position may limit purity [ 20 ]. However, further exploration in this area and better characterization of these systems fill a useful niche where a unique biomarker is required for distinguishing populations. Acoustic separation. The acoustic radiation force may be manipulated for density-based, equilibrium separation a , b [ 88 ] reproduced by permission of The Royal Society of Chemistry. As deduced from Eq.
These particles will be attracted to different parts of the channel: pressure nodes high density particles or antinodes low density particles. In an example of this, Petersson et al. RBCs were focused at the center line of the microfluidic channel node and lipids were focused near the side walls antinodes. An even simpler application of the acoustic standing wave is particle focusing for filtration [ 91 , 92 , 93 ]. Typically the focused particles, such as blood cells, exit through a centered outlet while clear fluid, or plasma-containing fluid, is collected from other outlets.
In principle, similar application of acoustic standing wave separation for particles with different compressibilities is possible. In flow segregation of polymeric particles based on compressibility in a millimeter-scale fluidic device has been demonstrated [ 94 ]; however, we have not found any report of cell separation based on this.
There have been several reports of particle sorting based on size [ 92 , 95 ]. The techniques that have been covered thus far may be considered equilibrium techniques where the equilibrium positions within a channel vary depending on cell or particle density.
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On the other hand, size-based acoustic standing wave separation techniques are kinetic. The acoustic radiation force acting on particles with different sizes but the same densities will have the same sign and therefore focus the particles to the same location in the channel. If a cell suspension is introduced near the walls of a primary channel and deflected toward the node at the center of a channel particles will cross streamlines at different rates.
As shown in Fig. Acoustic separation is being explored commercially for the binary separation of red blood cells and lipids ErySave AB, Sweden. This separation is expected to be used during surgery in adipose tissue where fat particles may cause complications. Other methods that perform the same function have numerous drawbacks which do not apply to this technology [ 89 ]. The abundance of tunable factors, all contributing to this resultant dielectrophoretic force field, allow for many types of DEP devices, which perform the sorting of heterogeneous populations of particles based on the intrinsic properties of 1 difference in polarizability between the particle and the surrounding medium and 2 the size of the suspended particle.
There exist advantages and disadvantages to AC and DC driving currents. Using an AC driving current enables the manipulation of the CM frequency dependence. DC current, on the other hand, allows for simplicity of device design by not requiring auxiliary equipment other than a voltage source. One significant disadvantage of DC current is the resulting electrochemical reactions on the electrode surface. These redox reactions result in a number of problems, most significantly convective flow, which can counteract particle trapping, and free radical generation that results in significant cellular damage [ 99 ].
The electrode geometry has been shown to be important, due to the effect on electric field non-uniformity, both through theoretical models and experimental results. Among others, Green et al. The magnitude and sign of the induced dipole moment, a function of the CM factor, is a crucial factor in design. If the complex permittivity of the particle is greater than that of the medium, then the CM factor is positive and the particle feels a force directed toward the field maximums, termed positive DEP pDEP , and vice versa negative, nDEP.
Particles exhibit positive CM factors in some ranges of frequency, and negative CM factors in other distinctly different frequency ranges. These types of DEP can be used to separate heterogeneous colloidal solutions based on where the crossover frequencies lie in frequency space. The imaginary component of the CM factor is acted upon by traveling waves, and will be explained in more detail in subsequent sections. DEP electrode setups are frequently fabricated as integrated aspects of a microfluidic channel.
The small length scales allow for highly defined non-uniform electric fields, thus more proficient use of the abilities of DEP. In many cases, the microfluidic flow of the DEP device is used only as a means to pass the particle-containing solution over the electrode surfaces. Gagnon et al. The long serpentine design was proposed to reduce faradic reactions on the electrode surface, avoiding electrochemical flow, electrolysis of water, and free radical generation.
In order to bring down the crossover frequency of the system, a zwitterionic small molecule 6-aminohexanioc acid was added to raise the complex permittivity of the medium, as shown previously by Zimmerman et al. Recently, exciting advances toward the creation of a truly integrated DEP microfluidic system have been made. By stringent design of the electrode geometry, a precise manipulation of DEP forces and fluid forces and their relation has been achieved.
DEP gates are an effective and creative method of selecting between multiple types of suspended particles entrained in fluid flow. Chen et al.
Experimentally, they also were able to accomplish separation between 4. Cheng et al. The design of the DEP gates were optimized based on average fluid velocity, taking advantage of a force balance between Stokes drag and DEP force. The combination of both label-free separation and trapping is an example of what can be accomplished by truly integrating DEP forces and fluid forces.
Altering the driving signal to the electrodes enables a new method of distinguishing between particles other than the crossover frequency. Recently, Cui et al. In this method, the frequency of the sinusoid wave is held constant, and particles can be selected based on size by changing the frequency of the square wave using a simple electrode design [ ].
This technique operates at a flow rate of only 0. The authors also suggest the ability to increase the flow rate with higher voltage and frequencies. One drawback to DEP chips is the slight difficulty in fabrication, especially aligning gate electrodes. Cummings et al.
Rather than patterning conductive electrodes on the device, they pattern insulating posts of varying designs in the channel. This allows for remote electrode placement, as the special non-uniformities in the electric field are resultant from the insulating boundaries, with operation down to frequencies that would normally cause detrimental reactions. In this form of DEP fluid flow is resultant from electroosmosis, which restricts the flow rate [ ]; however, this technique has demonstrated the ability to create ordered streams and separate particles from suspension.
Recently, Vahey et al. The device consists of an inlet diffusive mixer, common to microfluidic devices, coupled with a long straight channel with angled electrodes running the diagonal of the long channel. The diffusive mixer results in a gradient of conductivity across the short dimension of the channel spanning 9. This separation, based on differences in membrane permeabilites as in the case of live versus dead cells , is a feat thus far unmatched by many other techniques. These have shown that twDEP makes use of the imaginary component of the CM factor and, furthermore, that the real and imaginary components of the CM factor are acted upon independently by the electric field and correspond to trapping and translational forces, respectively.
This wealth of theoretical information has allowed successful creation of separation devices based on twDEP. Using this design, Cheng et al.
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These innovations of late have created new possibilities for the label-free separation and sorting abilities of DEP. The integration of microfluidic force fields and DEP fields has allowed for creative solutions for label-free applications. Presently, the state of these integrated, multi-module devices is promising. Separating based on electrical characteristics specifically the polarizability allows for distinguishing between live and dead cells, as well as types of bacteria by physical properties other than just size. In order for this technology to become truly useful and accessible, it is necessary to create large, thorough libraries of characteristic DEP data, such as crossover frequencies.
Some of the experiments have been done, among them an analysis of how gene expression profiles are affected by DEP [ ], characterization of the crossover frequencies of all mononuclear components of blood [ ], linking how biological states, such as RBC starvation and glutaraldehyde cross-linking, lead to changes in crossover frequencies [ ], and mapping lateral displacement as a function of particle size in specific general electrode designs [ ].
Optical techniques for manipulating particles have recently been developed into effective platforms for the sorting of heterogeneous populations.