Borys-Wójcik et al. Medical Journal of 2019 DOI: 10.2478/acb-2019-0002 Received: 06.03.2019 Accepted: 26.03.2019

Historical background of umbilical stem

Sylwia Borys-Wójcik1 2 3, Sandra Knap3, Wojciech 4 5 1, Bartosz Kempisty1,3,6 , Małgorzata Józkowiak , Katarzyna Stefańska Pieńkowski , Paweł Gutaj , Małgorzata Bruska Abstract Umbilical cord is a waste material, and therefore does not raise ethical concerns related to its use for rese- arch and medicine. Stem cells from umbilical cord have a significant advantage over cells from other sour- ces. First, the umbilical cord is an infinite source of stem cells, because it can be taken theoretically during each delivery. Secondly, acquisition of umbilical cord is a non-invasive, safe procedure for mother and child. Thirdly, the transplantation of umbilical cord stem cells is associated with a lower risk of infection and a less-frequent “graft versus host” reaction. In this work, the authors present a historical background of rese- arch on the cell from its discovery to modern times characterized by highly advanced methods of obtaining stem cells from umbilical cord and from other sources.

Running title: History of umbilical stem cell culture

Keywords: umbilical stem cells, history of umbilical stem cells

1Department of Anatomy, Poznan University of Medical Sciences, Poznan, Poland 2Department of Toxicology, Poznan University of Medical Sciences, Poznan, Poland 3Department of Histology and , Poznan University of Medical Sciences, Poznan, Poland 4Division of Perinatology and Women’s Diseases, Poznan University of Medical Sciences, Poznan, Poland 5Department of Reproduction, Poznan University of Medical Sciences, Poznan, Poland 6 * Correspondence: [email protected] FullDepartment list of author of Obstetrics information and is Gynecology, available at University the end of Hospital article and Masaryk University, Brno, Czech Republic 12 Borys-Wójcik et al. Medical Journal of Cell Biology (2019)

Introduction The Twentieth Century The construction of living organisms has been - the subject of people’s interests since antiquity. In gist and histologist, attempted to breed nucleated the available literature one can find information redIn blood 1903, cells. Justin He Marrie was also Jolly, referencing a French hematolo the work were making various theories about the structure thanks to the breeding medium he developed, in- ofthat organisms. in the years Anaxagoras 500-428 from BC, Greek Kladzom philosophers - an an- creasedof the Germanthe size of botanist the cultured Gotlieb cells, Haberland, but did not who ob- cient atomist claimed that reality is built from mat- tain their multiplication. The blood cells cultured by ter consisting of immutable and infinite “seeds” and from self-governing reason, which was supposed to they gradually lost ability to cell divisions until total set in motion and cause connection and disconnec- arrestJustin Jollyfew monthsdivided afterfor the the first start 15 ofdays, cultivation after which [7]. tion of elements [1]. Beebe - zoologist and oceanographer, and ocean- The Seventeenth to Nineteenth Centuries In 1906, two American scientists, Charles William In 1665, Robert Hooke, an English naturalist, ob- to in vitro maintain a dog lymphosarcoma cells. served the existence of compartments in tissues, Theographer breakthrough James Ewing, moment made in ain successful vitro cell attemptculture describing the cell - the smallest functional and is considered to have occurred in 1907 [8]. In this structural unit of living organisms capable of car- - and anatomist, achieved success in in vitro culti- man scholars: zoologist - Theodor Schwann and vationyear, Ross of diverse Granville nerve Harrison, cells ofan theAmerican radial biologistnerve of rying out all life processes [2]. In 1839, two Ger amphibian embryos [9]. Harrison placed fragments cellular theory that all living organisms are made of the nervous tissue of the frog’s embryo in a drop ofbotanist cells. The - Matthias above theory Jacob Schleidenwas created put in forwardtwo stag a- of congealed lymph. This experiment showed that axons of nerve cells are formed from protrusions plant organisms were made of cells, and a year later that grow out of the body of nerve cells. Harrison Theodores. In 1838 Schwann Mattihas extended Jacob Schlieden the application stated that of the all also observed the movement of the cytoplasm from - the cell body to the protrusions [10]. During the mal cell [3,4]. The essence of in vitro cell culture is first observations, Harrison used frog cells, as they toGerman maintain botanist their byvital stating functions the existence outside theof an living ani were not hard to maintain in cells cultures. The frog organism [5]. Interest in the culture of cells, tissues is a cold-blooded animal, so its cells do not require and organs outside the body became the subject of incubation under constant conditions. In addition, interest of scientists shortly after the discovery of tissue regeneration is more effective in lower verte- brates. Therefore, Harrison supposed that the expe- - rience would have a greater chance of success than iologytheir existence. and topographic Carl Friedrich anatomy Wilhelmin 1856 developed Ludwig - with the use of mammalian cells [11]. Ross Harri- techniquesGerman physician, of organ physiologist, perfusion after professor removing of physthem son is considered the creator of the methods used in from the body. The essence of the experiment was cell culture [1]. The American also developed a cell to pump blood through organs taken from the body. culture technique “in a hanging drop”. Then, thanks Three years later - in 1859, a French neurologist to cooperation with Montrose Thomas Burrows, a Edmé Félix Alfred Vulpian isolated fragments of surgeon and pathologist, he succeeded in develop- tadpole tail and attempted to breed them in water. ing a cell culture technique in a drop of clotted plas- ma placed on a microscope slide. Montorse Thom- their survival in water, did not reproduce. Howev- as Burrows began his collaboration with Harrison er,Cells the isolated experience by Edmé of the Félix French Alfred neurologist Vulpian, showeddespite that cell survival in vitro is possible [1]. Twenty-one Prize winner in physiology and medicine in 1912. years later, the English clinician Sidney Ringer con- thanks to Alexis Carrel, a French surgeon, Nobel ducted research on the work of the cardiac muscle by him to Harrison’s laboratory to learn about the of animals placed in 0.75% saline and the effect of Burrows, as an assistant to Alexis Carrel, was sent additional substances added to it (blood, albumin, Harrison’s techniques for research on warm-blood- potassium). The British researcher proved that it is edcell animals. culture techniques.Montorse Thomas Carell’s Burrows goal was has to adoptbeen possible to survive the organs for a long time, pro- very successful during his cooperation with Haris- vided they are kept in a sodium chloride solution son. In 1910 he successfully established a tissue - culture of embryonic chicken cells [1]. After return- man embryologist Wilhelm Roux proved that it is possiblewith small to amountskeep the ofnerve potassium. plate obtained In 1885, from the Ger the a number of improvements to Harrison’s culture chicken embryo in a heated saline solution. Unfor- techniques.ing, Thomas These Burrows improvements and Alexis Carrel included, introduced among tunately, all attempts carried out resulted in a bac- other things, the use of a lymph instead of blood terial or fungal infection [6]. plasma and the addition of an extract from the em- 13 Borys-Wójcik et al. Medical Journal of Cell Biology (2019) bryo to the nutrient medium used for . structed a device extremely helpful in conducting Improvements made by scientists have contributed long-term cell culture - the so-called roller tube. to, inter alia, improving the growth of tissue cul- From the late 1930s, the first reports on the thera- peutic use of umbilical cord blood were also made. with Burrows, successfully maintained cellular ex- About ten years later, antibiotics began to be used in tures. In the years 1911-1912 Carrel, in cooperation cell culture [12], and in 1948 the first synthetic cell cells became the subject of their further interest culture medium was developer [1]. [1]plants from cats, dogs, rats or guinea pigs. Cancer culture introducing a number of recommendations been a specific revolution in in vitro research. The . Alexis Carrel made a breakthrough in tissue The discovery of the HeLa tumor cell line has the collection of material and the establishment andfor aseptic maintenance culture of techniques. the culture Carrelshould assumed take place that in diagnosedHeLa line was with prepared cervical by cancer Elexis Carreland sent and Georgeto the accordance with the principles of antiseptics. The Gey. In 1951, tissues were removed from a patient place of operation should be a sterile box and the blood was then used as a cell culture medium. By tools used should be sterile. He also assumed the studyingTissue Culture the tumor Laboratory. cells obtained Human from umbilical the tumor cord the method of culture in a thrombus blood clot. He continuously and do not die. During the previous wasuse ofalso sterile the originator media. Alexis of cell Carrel culture also in aintroduced coagulum studies,(HeLa line), the Geycells noticed died after that a these few days cells of are culture dividing as formed by mixing embryo extract of chicken embry- os and plasma [12]. The experience of the French manyopposed scientific to the newly papers discovered have been HeLa published cells. Several on the of an embryonic extract and asepsis during cell cul- findingsstudies have [16]. been carried out using HeLa cells and tures,Nobel itPrize is possible winner to showed keep the that, cells thanks out of to the the body use - Maruyama took effective attempts to harvest stem lar and continuous passage of fibroblasts from the cellsMore from than the umbilical a decade vein. later, Maruyama a Japanese proposed scholar, ob- heartfor many of the years. chicken Alexis to the Carrel, fresh thanks breeding to themedium, regu taining stem cells by cannulation of the umbilical vein - and by means of enzymatic activity of trypsin [17]. ture was continued for the next thirty-four years by The seventies of the twentieth century were a Albertobtained H. Ebeling a continuous - using culture. it to test Carrela’s germicides cell [13] cul. time of increased attempts to obtain pluripotent cells. In 1981, mouse pluripotent embryonic stem cell lines with normal karyotype were successful- Carrela’s close associate was also Charles Augustus ly isolated from blastocysts [18]. Seven years lat- andLinbergh Europe. - an PilotAmerican was theairman originator who became and contrac famous- er, in 1988, on the initiative of American doctors: for the first lonely passage between North America tor of a large number of devices used by Carrela for firstHana trial Broxmeyer in the history and Joanna of umbilical Krutzberg cord blood and prof. cell workedtissue culture. on the Lidbergh construction constructed, of instruments among usedothers, in transplantationEliane Gluckman [19]. at the Umbilical Paris Clinic cord undertookblood is a richthe thea press research for crushing [14]. chicken embryos. Carrel also source of pluripotent cells that can be dedifferen- Another breakthrough in the subject of in vitro tiated into any type of cell in the human body. An cell culture took place in 1916, when Francis Peyton umbilical cord blood of a newborn sister was given to the boy who was ill with Fanconi’s anemia. A few days after the transplantation, blood cells derived aRous proteolytic - American enzyme physician - trypsin. and Tryspin pathologist, was used Nobel to from the donor cells were found in his bloodstream. dissociatelaureate in single 1966 withcells Jonesfrom tissuefor the fragments. first time use After of The first allogeneic transplantation of frozen and dissociation, the cells were placed in the culture banked umbilical cord blood was performed in medium, resulting in a suspension of viable cells. 1993. A few years later, it was proved that the cord The idea of using trypsin to dissociate individual blood has enough stem cells to repopulate the recip- cells is practiced to this day [15]. ient’s marrow. Evidence has also been provided that The 1930s was the period of further develop- there is less risk of allograft rejection compared to ment of scientific in vitro cell culture techniques. bone marrow transplantation [20]. The first cord blood stem cell transplant in Poland took place in University is considered as the forerunner of tissue 1994 to treat a boy suffering from acute myeloge- Professor Zygmunt Grudzinski of the Jagiellonian- nous leukemia. feller Foundation scholarship holder, completed an culture in Poland. Professor Grudzinski, as a Rocke- The Twenty-First Century la - professor Maxirnov. The Pole had the opportu- In 2001, more than one blood unit was trans- nityinternship to learn in thethe methodslaboratory of ofchicken the student embryo of Carretissue planted in the USA. In Poland, a similar transplant was performed in a patient with refractory chemo- - therapy-resistant acute leukemia in 2003. In March culture in a hanging drop. In 1933, Alexis Carrel, in cooperation with cell biologist Georg Otto Gey, con 14 Borys-Wójcik et al. Medical Journal of Cell Biology (2019)

2007, the first Polish transplant of banked cord 16. Jones HW, McKusick VA, Harper PS, Wuu KD. George Otto Gey. (1899- - 17. 1970). The HeLa cell and a reappraisal of its origin. Obstet Gynecol. lectedblood wasin 2004 performed [21]. at the Children’s Hematology 1971;38:945–9. Jiménez N, Krouwer VJD, Post JA. A new, rapid and reproducible method Clinic in Wroclaw. The blood for transplant was col 18. to obtain high quality endothelium in vitro. Cytotechnology. 2013;65:1– Conclusion 14; DOI:10.1007/s10616-012-9459-9. 19. Evans MJ, Kaufman MH. Establishment in culture of pluripotential cells- Stem cell transplants taken from the umbilical from mouse embryos. Nature. 1981;292:154–6. cord over the years gain a wider range of applica- DOI:10.1182/blood-2013-02-453175.Ballen KK, Gluckman E, Broxmeyer HE. Umbilical cord blood trans tions and the use of cells stored in banks contributes 20. Bielecplantation: B, Stojko the R. first Stem 25 cells years of umbilical and beyond. blood Blood. cord - 2013;122:491–8; therapeutic use. to saving or improving the quality of life of many pa- 21. Postepy Hig Med Doswiadczalnej Online. 2015;69:853–63. since 2011, more than 1194 umbilical cord stem cell DOI:10.5604/17322693.1143052.Piskorska-Jasiulewicz MM, Witkowska-Zimny M. Perinatal sources of transplantstients. The data were of conducted Polish Stem in PolandCell Bank [22]. show that, 22. stem cells. Postepy Hig Med Doswiadczalnej Online. 2015;69:327–34; www.pbkm.pl/o-komorkach-macierzystych/gdzie-wykorzystuje-sie- Macierzystych-komorki-macierzyste/przeszczepienia-pbkm PBK. Lista przeszczepień PBKM (accessed - PBKM May n.d. 13, https:// 2019). Ethical approval The conducted research is not related to either hu- man or animal use.

Acknowledgements This publication and its results are an outcome of a cooperation and Polish Ministry of Science and Higher Education, with Insti- between Poznan University of Medical Sciences (Poznań, Poland) the “Professional PhD” programme. tute of Advanced Sciences Sp. z o.o. (Poznań, Poland), as a part of Corresponding author

Fax:Małgorzata +48 61 Bruska,8546565, Department e-mail: [email protected]. of Anatomy, Poznan University of Medical Sciences, 6 Ś�więcickiego St., 60-781 Poznań, Poland Tel./ Conflict of interest statement The authors declare they have no conflict of interest.

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