60:
192:
digestion of the extracellular matrix will not be complete. After the enzymes have been removed from the tissue by perfusing it with a second solution that does not contain enzymes, cells can be mechanically separated or dissociated. A simple technique for dissociating cells involves cutting the tissue into small chunk before agitating the chunks in a solution using a pipette.
84:, during which an automated analyser inspects a narrow stream of cells. In one version of this technique, a light is shone on the stream of cells, and the analyser detects the reflected light or fluorescence before using this information to rapidly manoeuvre the cells of interest into a collection chamber.
191:
These enzymes, in addition to digesting the extracellular matrix, can also digest other important proteins essential for the cells of interest to function. If cells are exposed to these enzymes for too long then cell death results, but if they are not exposed to the enzymes for long enough then
79:
The techniques required to obtain isolated cells vary depending on the cell type required. Circulating cells such as blood cells or some tumour cells can be isolated by taking a blood sample. As blood samples contain a mixture of many different cell types, a method of separating out cells into
112:
After a tissue specimen has been obtained, it must be surrounded or perfused by a solution at an appropriate temperature containing the salts and nutrients required to keep the cells alive. This may be performed by simply submerging the tissue in the solution, or may involve more complex
997:
Kloskowski T, Uzarska M, Gurtowska N, Olkowska J, Joachimiak R, Bajek A, Gagat M, Grzanka A, Bodnar M, Marszałek A, Drewa T (April 2014). "How to isolate urothelial cells? Comparison of four different methods and literature review".
491:
Kloskowski T, Uzarska M, Gurtowska N, Olkowska J, Joachimiak R, Bajek A, Gagat M, Grzanka A, Bodnar M, Marszałek A, Drewa T (April 2014). "How to isolate urothelial cells? Comparison of four different methods and literature review".
39:. After the matrix proteins have been digested, cells remain loosely bound together but can be gently separated mechanically. Following isolation, experiments can be performed on these single isolated cells including
161:. Isolation of cells from some tissues may be improved by oxygenating the solution. In the initial stages, perfusing the tissue with a solution that does not contain calcium is useful particularly when isolating
92:
When dealing with solid tissues, obtaining tissue for cell isolation may be more challenging. Surplus human tissue can sometimes be obtained at the time of planned surgery, for example specimens of
200:
Isolated cells can be used to study how cells work, how they change in response to disease, and how they are affected by drugs. An example of an experimental technique which uses isolated cells is
224:
Cell isolation can also be used as part of a treatment. Isolation of pancreatic islet cells, followed by their subsequent culture and transplantation, has been used to treat patients with
153:. The concentrations of these electrolytes must be carefully balanced, paying attention to osmotic pressure. The acidity of the solution must be regulated, often using a
456:
Yates JC, Dhalla NS (February 1975). "Structural and functional changes associated with failure and recovery of hearts after perfusion with Ca2+-free medium".
27:), other cell types that are found in solid tissue require specific techniques to separate them into individual cells. This may be performed by using
105:. Other tissues such as samples of pancreas or bladder may be taken as a biopsy. Alternatively, tissue from animals is frequently obtained by
217:
which uses antibodies tagged with a fluorescent marker to identify where proteins are located within a cell. Isolated cells can also be used for
934:
818:
584:
Frank, J.; Biesalski, H. K.; Dominici, S.; Pompella, A. (January 2000). "The visualization of oxidant stress in tissues and isolated cells".
273:
633:
955:"Finding degrees of separation: experimental approaches for astroglial and oligodendroglial cell isolation and genetic targeting"
537:"Finding degrees of separation: experimental approaches for astroglial and oligodendroglial cell isolation and genetic targeting"
23:
from a solid block of tissue or cell suspension. While some types of cell naturally exist in a separated form (for example
1050:
763:
Ricordi C, Lacy PE, Finke EH, Olack BJ, Scharp DW (April 1988). "Automated method for isolation of human pancreatic islets".
184:) are often used when isolating cells from the heart or bladder. General-purpose enzymes that digest many sorts of protein (
102:
188:) may also be used. When isolating cells from brain tissue, other enzymes that break down DNA (DNAases) may be required.
122:
917:
Bhagwat N, Carpenter EL (2017). "Flow
Cytometric Methods for Circulating Tumor Cell Isolation and Molecular Analysis".
256:
Bhagwat N, Carpenter EL (2017). "Flow
Cytometric Methods for Circulating Tumor Cell Isolation and Molecular Analysis".
225:
213:
using dyes that emit light in response to calcium to measure how calcium is regulated within the cell, and
839:"Mobilization, Isolation and Characterization of Stem Cells from Peripheral Blood: a Systematic Review"
118:
36:
214:
162:
68:
64:
48:
1023:
788:
517:
801:
Rector K, Liu Y, Van Zant G (2013). "Comprehensive
Hematopoietic Stem Cell Isolation Methods".
1015:
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166:
114:
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94:
81:
20:
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237:
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921:. Advances in Experimental Medicine and Biology. Vol. 994. pp. 105–118.
260:. Advances in Experimental Medicine and Biology. Vol. 994. pp. 105–118.
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684:
424:
377:
360:
265:
202:
181:
40:
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80:
different types must be used. The most commonly used method for this is
24:
605:
315:
142:
138:
130:
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513:
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283:
784:
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185:
177:
146:
125:
These solutions contain precise concentrations of electrolytes including
597:
150:
134:
32:
646:
173:
126:
28:
881:
Shang L, Hosseini M, Liu X, Kisseleva T, Brenner DA (January 2018).
627:
Kieffer TJ, Woltjen K, Osafune K, Yabe D, Inagaki N (October 2017).
919:
Isolation and
Molecular Characterization of Circulating Tumor Cells
258:
Isolation and
Molecular Characterization of Circulating Tumor Cells
685:"Methods for isolating atrial cells from large mammals and humans"
361:"Methods for isolating atrial cells from large mammals and humans"
158:
221:, in which a single cell multiplies to create a colony of cells.
721:"Methods in cardiomyocyte isolation, culture, and gene transfer"
409:"Methods in cardiomyocyte isolation, culture, and gene transfer"
805:. Methods in Molecular Biology. Vol. 976. pp. 1–15.
176:
can then be added to the solution. Enzymes that digest
883:"Human hepatic stellate cell isolation and characterization"
683:
Voigt N, Pearman CM, Dobrev D, Dibb KM (September 2015).
359:
Voigt N, Pearman CM, Dobrev D, Dibb KM (September 2015).
205:, used to study how charged particles flow across the
165:, as the absence of calcium causes separation of the
117:. Commonly used solutions included modifications of
719:Louch WE, Sheehan KA, Wolska BM (September 2011).
407:Louch WE, Sheehan KA, Wolska BM (September 2011).
953:Chew LJ, DeBoy CA, Senatorov VV (October 2014).
535:Chew LJ, DeBoy CA, Senatorov VV (October 2014).
629:"Beta-cell replacement strategies for diabetes"
19:is the process of separating individual living
8:
843:Journal of Oral & Maxillofacial Research
725:Journal of Molecular and Cellular Cardiology
689:Journal of Molecular and Cellular Cardiology
458:Journal of Molecular and Cellular Cardiology
413:Journal of Molecular and Cellular Cardiology
402:
400:
398:
396:
365:Journal of Molecular and Cellular Cardiology
354:
352:
350:
348:
346:
344:
303:Frontiers in Cell and Developmental Biology
35:that binds these cells together within the
837:Yosupov N, Haimov H, Juodzbalys G (2017).
978:
898:
862:
744:
717:Isolation of Ventricular Cardiomyocytes:
700:
654:
560:
432:
376:
324:
314:
209:. Complementary techniques include
58:
915:Isolation of Circulating tumour cells:
799:Isolation of Haematopoetic stem cells:
248:
97:are often excised and discarded during
879:Isolation of Hepatic stellate cells:
835:Isolation of Mesenchymal stem cells:
761:Isolation of Pancreatic Islet Cells:
297:Hu P, Zhang W, Xin H, Deng G (2016).
7:
681:Isolation of Atrial Cardiomyocytes:
299:"Single Cell Isolation and Analysis"
123:Krebs and Henseleit's solution
14:
634:Journal of Diabetes Investigation
995:Isolation of Urothelial cells:
959:Journal of Neuroscience Methods
951:Isolation of Neuroglial cells:
541:Journal of Neuroscience Methods
971:10.1016/j.jneumeth.2014.08.017
553:10.1016/j.jneumeth.2014.08.017
103:coronary artery bypass surgery
1:
203:patch clamp electrophysiology
41:patch clamp electrophysiology
586:Histology and Histopathology
470:10.1016/0022-2828(75)90011-5
211:calcium fluorescence imaging
45:calcium fluorescence imaging
927:10.1007/978-3-319-55947-6_5
887:Journal of Gastroenterology
811:10.1007/978-1-62703-317-6_1
737:10.1016/j.yjmcc.2011.06.012
702:10.1016/j.yjmcc.2015.07.006
425:10.1016/j.yjmcc.2011.06.012
378:10.1016/j.yjmcc.2015.07.006
266:10.1007/978-3-319-55947-6_5
1067:
1012:10.1007/s13577-013-0070-y
900:10.1007/s00535-017-1404-4
506:10.1007/s13577-013-0070-y
316:10.3389/fcell.2016.00116
63:Image of isolated adult
855:10.5037/jomr.2017.8101
95:right atrial appendage
71:
1051:Laboratory techniques
777:10.2337/diab.37.4.413
115:Langendorff perfusion
113:arrangements such as
62:
803:Stem Cells and Aging
37:extracellular matrix
215:immunocytochemistry
69:confocal microscopy
49:immunocytochemistry
598:10.14670/HH-15.173
167:intercalated disks
99:open heart surgery
72:
936:978-3-319-55946-9
820:978-1-62703-316-9
647:10.1111/jdi.12758
275:978-3-319-55946-9
75:Circulating cells
1058:
1031:
992:
982:
948:
912:
902:
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404:
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339:
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328:
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288:
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253:
163:cardiac myocytes
65:cardiac myocytes
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1065:
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675:Further reading
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226:Type 1 Diabetes
198:
90:
77:
57:
12:
11:
5:
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1038:
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949:
935:
913:
877:
833:
819:
797:
759:
715:
678:
676:
673:
671:
670:
641:(3): 457–463.
619:
592:(1): 173–184.
576:
527:
483:
448:
392:
340:
289:
274:
247:
245:
242:
241:
240:
233:
230:
197:
194:
89:
86:
82:flow cytometry
76:
73:
56:
53:
31:to digest the
17:Cell isolation
13:
10:
9:
6:
4:
3:
2:
1063:
1052:
1049:
1047:
1044:
1043:
1041:
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1021:
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1005:
1001:
994:
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986:
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865:
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848:
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830:
826:
822:
816:
812:
808:
804:
798:
794:
790:
786:
782:
778:
774:
771:(4): 413–20.
770:
766:
760:
756:
752:
747:
742:
738:
734:
731:(3): 288–98.
730:
726:
722:
716:
712:
708:
703:
698:
694:
690:
686:
680:
679:
674:
666:
662:
657:
652:
648:
644:
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635:
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623:
620:
615:
611:
607:
603:
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563:
558:
554:
550:
546:
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538:
531:
528:
523:
519:
515:
511:
507:
503:
499:
495:
487:
484:
479:
475:
471:
467:
464:(2): 91–103.
463:
459:
452:
449:
444:
440:
435:
430:
426:
422:
419:(3): 288–98.
418:
414:
410:
403:
401:
399:
397:
393:
388:
384:
379:
374:
370:
366:
362:
355:
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207:cell membrane
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88:Solid tissues
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66:
61:
54:
52:
50:
46:
42:
38:
34:
30:
26:
22:
18:
1006:(2): 85–93.
1003:
999:
962:
958:
918:
890:
886:
846:
842:
802:
768:
764:
728:
724:
692:
688:
638:
632:
622:
589:
585:
579:
544:
540:
530:
500:(2): 85–93.
497:
493:
486:
461:
457:
451:
416:
412:
368:
364:
306:
302:
292:
257:
251:
238:Cell sorting
223:
219:cell culture
199:
190:
182:collagenases
172:Proteolytic
171:
121:solution or
111:
109:the animal.
91:
78:
67:taken using
16:
15:
893:(1): 6–17.
107:sacrificing
25:blood cells
1040:Categories
1000:Human Cell
965:: 125–47.
695:: 187–98.
547:: 125–47.
494:Human Cell
371:: 187–98.
244:References
55:Techniques
849:(1): e1.
606:0213-3911
186:proteases
155:pH buffer
143:phosphate
139:magnesium
131:potassium
1020:24368576
989:25169049
945:28560670
909:29094206
873:28496961
829:23400430
793:20974701
765:Diabetes
755:21723873
711:26186893
665:28984038
614:10668208
571:25169049
514:24368576
443:21723873
387:26186893
335:27826548
284:28560670
232:See also
178:collagen
157:such as
147:chloride
119:Tyrode's
101:such as
51:.
33:proteins
1028:9966872
980:4171043
864:5423306
785:3288530
746:3164875
656:5934267
562:4171043
522:9966872
478:1121035
434:3164875
326:5078503
309:: 116.
174:enzymes
151:glucose
135:calcium
29:enzymes
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323:
282:
272:
149:, and
127:sodium
47:, and
1046:Cells
1024:S2CID
789:S2CID
518:S2CID
159:HEPES
21:cells
1016:PMID
985:PMID
941:PMID
931:ISBN
905:PMID
869:PMID
825:PMID
815:ISBN
781:PMID
751:PMID
707:PMID
661:PMID
610:PMID
602:ISSN
567:PMID
510:PMID
474:PMID
439:PMID
383:PMID
331:PMID
280:PMID
270:ISBN
196:Uses
1008:doi
975:PMC
967:doi
963:236
923:doi
895:doi
859:PMC
851:doi
807:doi
773:doi
741:PMC
733:doi
697:doi
651:PMC
643:doi
594:doi
557:PMC
549:doi
545:236
502:doi
466:doi
429:PMC
421:doi
373:doi
321:PMC
311:doi
262:doi
1042::
1022:.
1014:.
1004:27
1002:.
983:.
973:.
961:.
957:.
939:.
929:.
903:.
891:53
889:.
885:.
867:.
857:.
845:.
841:.
823:.
813:.
787:.
779:.
769:37
767:.
749:.
739:.
729:51
727:.
723:.
705:.
693:86
691:.
687:.
659:.
649:.
637:.
631:.
608:.
600:.
590:15
588:.
565:.
555:.
543:.
539:.
516:.
508:.
498:27
496:.
472:.
460:.
437:.
427:.
417:51
415:.
411:.
395:^
381:.
369:86
367:.
363:.
343:^
329:.
319:.
305:.
301:.
278:.
268:.
228:.
169:.
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969::
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925::
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897::
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853::
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809::
795:.
775::
757:.
735::
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699::
667:.
645::
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596::
573:.
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524:.
504::
480:.
468::
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423::
389:.
375::
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313::
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286:.
264::
180:(
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