vtioВестник трансплантологии и искусственных органовRussian Journal of Transplantology and Artificial Organs1995-1191Academician V.I.Shumakov National Medical Research Center of Transplantology and Artificial Organs", Ministry of Health of the Russian Federation10.15825/1995-1191-2022-1-48-55vtio-1441Research ArticleРегенеративная медицина и клеточные технологииRegenerative Medicine and Cell TechnologiesРазработка подходов к бесферментному получению островковой ткани из поджелудочной железыDevelopment of approaches to enzyme-free isolation of pancreatic isletsСкалецкаяГ. Н.SkaletskayaG. N.

Скалецкая Галина Николаевна

23182, Москва, ул. Щукинская, д. 1.

Тел.: (499) 190-42-66; (903) 771-18-13

Galina Skaletskaya

1, Shchukinskaya str., Moscow, 123182

Phone: (499) 190-42-66; (903) 771-18-13

skalink@mail.ruСкалецкийН. Н.SkaletskiyN. N.

Москва

Moscow

БубенцоваГ. Н.BubentsovaG. N.

Москва

Moscow

СевастьяновВ. И.SevastianovV. I.

Москва

Moscow

ФГБУ «Национальный медицинский исследовательский центр трансплантологии и искусственных органов имени академика В.И. Шумакова» Минздрава РоссииРоссияShumakov National Medical Research Center of Transplantology and Artificial OrgansRussian Federation2022020220222414855Copyright © Скалецкая Г.Н., Скалецкий Н.Н., Бубенцова Г.Н., Севастьянов В.И., 20222022Скалецкая Г.Н., Скалецкий Н.Н., Бубенцова Г.Н., Севастьянов В.И.Skaletskaya G.N., Skaletskiy N.N., Bubentsova G.N., Sevastianov V.I.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.transpl.ru/vtio/article/view/1441

Успешность аллотрансплантации островков поджелудочной железы при лечении пациентов с трудноуправляемым течением сахарного диабета 1-го типа зависит главным образом от количества и качества островков, изолируемых из поджелудочной железы посмертных доноров с помощью ферментных препаратов, прежде всего коллагеназы. Многочисленные исследования по совершенствованию и стандартизации методов выделения островков достигли в последнее десятилетие предела своих возможностей, что сделало невозможным дальнейшее увеличение количества и качества клинических трансплантаций. Учитывая негативное влияние применения коллагеназной техники на морфофункциональные свойства изолируемых островков, в настоящей работе изучена возможность бесферментного получения островковой ткани, очищенной от экзокринного балласта. Эксперименты, проведенные с использованием поджелудочной железы новорожденных и молодых кроликов, показали реальность разработки методических подходов к получению островковоподобных культур без применения экзогенных ферментов.

The success of pancreatic islet allotransplantation in the treatment of patients with a difficult-to-manage type 1 diabetes depends mainly on the quantity and quality of islets isolated from the pancreas of deceased donors using enzyme preparations, primarily collagenase. Numerous studies on improvement and standardization of islet isolation techniques have reached their limits in the last decade. This has made it impossible to further boost the number and quality of clinical transplants. Taking into account the negative impact of collagenase technique on the morphofunctional properties of isolated islets, this work has studied the possibility of enzyme-free isolation of islet tissue purified of exocrine ballast. Experiments using the pancreas of newborn and young rabbits showed that developing methodological approaches to obtaining islet-like cultures without the use of exogenous enzymes is feasible.

поджелудочная железаэкзокринная тканьостровкиизоляцияколлагеназакроликифлотирующие островковоподобные культурыpancreasexocrine tissueisletsisolationcollagenaserabbitsfloating islet-like culturesReferencesShapiro AM, Lakey JR, Ryan EA, Korbutt GS, Toth E, Warnock GL et al. Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen. N Engl J Med. 2000; 343: 230–238.Shapiro AM, Lakey JR, Ryan EA, Korbutt GS, Toth E, Warnock GL et al. Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen. N Engl J Med. 2000; 343: 230–238.Barton FB, Rickels MR, Alejandro R, Hering BJ, Wease S, Naziruddin B et al. Improvement in outcomes of clinical islet transplantation: 1999–2010. Diabetes Care. 2012; 35: 1436–1445.Barton FB, Rickels MR, Alejandro R, Hering BJ, Wease S, Naziruddin B et al. Improvement in outcomes of clinical islet transplantation: 1999–2010. Diabetes Care. 2012; 35: 1436–1445.Misawa R, Ricordi C, Miki A, Barker S, Molano RD, Khan A et al. Evaluation of viable beta-cell mass is useful for selecting collagenase for human islet isolation: comparison of collagenase NB1 and liberase HI. Cell Transplant. 2012; 21: 39–47.Misawa R, Ricordi C, Miki A, Barker S, Molano RD, Khan A et al. Evaluation of viable beta-cell mass is useful for selecting collagenase for human islet isolation: comparison of collagenase NB1 and liberase HI. Cell Transplant. 2012; 21: 39–47.Linetsky E, Bottino R, Lehmann R, Alejandro R, Inverardi L, Ricordi C. Improved human islet isolation using a new enzyme blend, liberase. Diabetes. 1997; 46: 1120– 1123.Linetsky E, Bottino R, Lehmann R, Alejandro R, Inverardi L, Ricordi C. Improved human islet isolation using a new enzyme blend, liberase. Diabetes. 1997; 46: 1120– 1123.Alejandro R, Barton FB, Hering BJ, Wease S. 2008 Update from the Collaborative Islet Transplant Registry. Transplantation. 2008; 86: 1783–1788.Alejandro R, Barton FB, Hering BJ, Wease S. 2008 Update from the Collaborative Islet Transplant Registry. Transplantation. 2008; 86: 1783–1788.Sabek OM, Cowan P, Fraga DW, Gaber AO. The effect of isolation methods and the use of different enzymes on islet yield and in vivo function. Cell Transplant. 2008; 17: 785–792.Sabek OM, Cowan P, Fraga DW, Gaber AO. The effect of isolation methods and the use of different enzymes on islet yield and in vivo function. Cell Transplant. 2008; 17: 785–792.Bertuzzi F, Cainarca S, Marzorati S, Bachi A, Antonioli B, Nano R et al. Collagenase isoforms for pancreas digestion. Cell Transplant. 2009; 18: 203–206.Bertuzzi F, Cainarca S, Marzorati S, Bachi A, Antonioli B, Nano R et al. Collagenase isoforms for pancreas digestion. Cell Transplant. 2009; 18: 203–206.Szot GL, Lee MR, Tavakol MM, Lang J, Dekovic F, Kerlan RK et al. Successful clinical islet isolation using a GMP-manufactured collagenase and neutral protease. Transplantation. 2009; 88: 753–756.Szot GL, Lee MR, Tavakol MM, Lang J, Dekovic F, Kerlan RK et al. Successful clinical islet isolation using a GMP-manufactured collagenase and neutral protease. Transplantation. 2009; 88: 753–756.Wang Y, Paushter D, Wang S, Barbaro B, Harvat T, Danielson K et al. Highly purified versus filtered crude collagenase: comparable human islet isolation outcomes. Cell Transplant. 2011; 20: 1817–1825. PMID: 21396158.Wang Y, Paushter D, Wang S, Barbaro B, Harvat T, Danielson K et al. Highly purified versus filtered crude collagenase: comparable human islet isolation outcomes. Cell Transplant. 2011; 20: 1817–1825. PMID: 21396158.Kayton S, Poffenberger G, Henske J, Dai Ch, Thompson C, Aramandla R et al. Human islet preparations distributed for research exhibit a variety of insulin-secretory profiles. Am J Physiol Endocrinol Metab. 2015 Apr 1; 308 (7): E592–E602. doi: 10.1152/ajpendo.00437.2014.Kayton S, Poffenberger G, Henske J, Dai Ch, Thompson C, Aramandla R et al. Human islet preparations distributed for research exhibit a variety of insulin-secretory profiles. Am J Physiol Endocrinol Metab. 2015 Apr 1; 308 (7): E592–E602. doi: 10.1152/ajpendo.00437.2014.Rheinheimer J, Klarmann Ziegelmann P, Carlessi R, Ross Reck L, Bauer AC, Leitão B. Different digestion enzymes used for human pancreatic islet isolation: A mixed treatment comparison (MTC) meta-analysis. Islets. 2014; 6 (4): e977118. Published online 2014 Nov 7.Rheinheimer J, Klarmann Ziegelmann P, Carlessi R, Ross Reck L, Bauer AC, Leitão B. Different digestion enzymes used for human pancreatic islet isolation: A mixed treatment comparison (MTC) meta-analysis. Islets. 2014; 6 (4): e977118. Published online 2014 Nov 7.Khatri R, Hussmann B, Rawat D, Gürol AO, Linn T. Intraportal Transplantation of Pancreatic Islets in Mouse Model. J Vis Exp. 2018 May 5; (135): 57559. doi: 10.3791/57559.Khatri R, Hussmann B, Rawat D, Gürol AO, Linn T. Intraportal Transplantation of Pancreatic Islets in Mouse Model. J Vis Exp. 2018 May 5; (135): 57559. doi: 10.3791/57559.Saliba Y, Farès N. Isolation, Purification, and Culture of Mouse Pancreatic Islets of Langerhans. Methods Mol Biol. 2019; 1940: 255–265. doi: 10.1007/978-1-4939- 9086-3_18. PMID: 30788831.Saliba Y, Farès N. Isolation, Purification, and Culture of Mouse Pancreatic Islets of Langerhans. Methods Mol Biol. 2019; 1940: 255–265. doi: 10.1007/978-1-4939- 9086-3_18. PMID: 30788831.Corbin KL, West HL, Brodsky S, Whitticar NB, Koch WJ, Nunemaker CS. A Practical Guide to Rodent Islet Isolation and Assessment Revisited. Biol Proced Online. 2021 Mar 1; 23 (1): 7. doi: 10.1186/s12575-021-00143-x. PMID: 33641671.Corbin KL, West HL, Brodsky S, Whitticar NB, Koch WJ, Nunemaker CS. A Practical Guide to Rodent Islet Isolation and Assessment Revisited. Biol Proced Online. 2021 Mar 1; 23 (1): 7. doi: 10.1186/s12575-021-00143-x. PMID: 33641671.Dietrich I, Girdlestone J, Giele H. Differential cytokine expression in direct and indirect co-culture of islets and mesenchymal stromal cells. Cytokine. 2021 Dec 17; 150: 155779. doi: 10.1016/j.cyto.2021.155779.Dietrich I, Girdlestone J, Giele H. Differential cytokine expression in direct and indirect co-culture of islets and mesenchymal stromal cells. Cytokine. 2021 Dec 17; 150: 155779. doi: 10.1016/j.cyto.2021.155779.Hubber EL, Rackham CL, Jones PM. Protecting islet functional viability using mesenchymal stromal cells. Stem Cells Transl Med. 2021 May; 10 (5): 674–680. doi: 10.1002/sctm.20-0466.Hubber EL, Rackham CL, Jones PM. Protecting islet functional viability using mesenchymal stromal cells. Stem Cells Transl Med. 2021 May; 10 (5): 674–680. doi: 10.1002/sctm.20-0466.Brandhorst D, Brandhorst H, Johnson PRV. Enzyme Development for Human Islet Isolation: Five Decades of Progress or Stagnation? Rev Diabet Stud. 2017 Spring; 14 (1): 22–38.Brandhorst D, Brandhorst H, Johnson PRV. Enzyme Development for Human Islet Isolation: Five Decades of Progress or Stagnation? Rev Diabet Stud. 2017 Spring; 14 (1): 22–38.Gorelick FS, Otani T. Mechanisms of intracellular zymogen activation. Baillieres Best Pract Res Clin Gastroenterol. 1999; 13 (2): 227–240.Gorelick FS, Otani T. Mechanisms of intracellular zymogen activation. Baillieres Best Pract Res Clin Gastroenterol. 1999; 13 (2): 227–240.Piton G, Barbot O, Manzon C, Moronval F, Patry C, Navellou JC et al. Acute ischemic pancreatitis following cardiac arrest: a case report. JOP. 2010; 11 (5): 456–459.Piton G, Barbot O, Manzon C, Moronval F, Patry C, Navellou JC et al. Acute ischemic pancreatitis following cardiac arrest: a case report. JOP. 2010; 11 (5): 456–459.Dembinski A, Warzecha Z, Ceranowicz P, Tomaszewska R, Dembinski M, Pabianczyk M et al. Ischemic preconditioning reduces the severity of ischemia/reperfusion-induced pancreatitis. Eur J Pharmacol. 2003; 473 (2–3): 207–216.Dembinski A, Warzecha Z, Ceranowicz P, Tomaszewska R, Dembinski M, Pabianczyk M et al. Ischemic preconditioning reduces the severity of ischemia/reperfusion-induced pancreatitis. Eur J Pharmacol. 2003; 473 (2–3): 207–216.Trimble ER. In: Lanza RP, Chick WL. Pancreatic islet transplantation. 1994. Pancreatic islet-acinar relationships; 19–25.Trimble ER. In: Lanza RP, Chick WL. Pancreatic islet transplantation. 1994. Pancreatic islet-acinar relationships; 19–25.van Deijnen JH, Hulstaert CE, Wolters GH, van Schilfgaarde R. Significance of the peri-insular extracellular matrix for islet isolation from the pancreas of rat, dog, pig, and man. Cell Tissue Res. 1992; 267 (1): 139–146.van Deijnen JH, Hulstaert CE, Wolters GH, van Schilfgaarde R. Significance of the peri-insular extracellular matrix for islet isolation from the pancreas of rat, dog, pig, and man. Cell Tissue Res. 1992; 267 (1): 139–146.van Suylichem PT, van Deijnen JE, Wolters GH, van Schilfgaarde R. Amount and distribution of collagen in pancreatic tissue of different species in the perspective of islet isolation procedures. Cell Transplant. 1995; 4 (6): 609–614.van Suylichem PT, van Deijnen JE, Wolters GH, van Schilfgaarde R. Amount and distribution of collagen in pancreatic tissue of different species in the perspective of islet isolation procedures. Cell Transplant. 1995; 4 (6): 609–614.Otonkoski T, Banerjee M, Korsgren O, Thornell LE, Virtanen I. Unique basement membrane structure of human pancreatic islets: implications for beta-cell growth and differentiation. Diabetes Obes Metab. 2008; 10 (Suppl): 119–127.Otonkoski T, Banerjee M, Korsgren O, Thornell LE, Virtanen I. Unique basement membrane structure of human pancreatic islets: implications for beta-cell growth and differentiation. Diabetes Obes Metab. 2008; 10 (Suppl): 119–127.Jiang FX, Naselli G, Harrison LC. Distinct distribution of laminin and its integrin receptors in the pancreas. J Histochem Cytochem. 2002; 50 (12): 1625–1632.Jiang FX, Naselli G, Harrison LC. Distinct distribution of laminin and its integrin receptors in the pancreas. J Histochem Cytochem. 2002; 50 (12): 1625–1632.

The authors declare that there are no conflicts of interest present.