Islet isolation from the pancreas after pancreatectomy. Indications, methodology, evaluation of results

Aim. To develop and implement a method for isolating the islets of Langerhans from pancreatic tissue after pancreatectomy with islet autotransplantation.

Materials and Methods. The study used ten Wiesenau miniature pig pancreases, 30 Wistar rat pancreases, eight resected human pancreases, and five human pancreases from extensive resection or pancreatectomy.

Results. Islets of Langerhans completely devoid of human and experimental animals’ acinar tissue were obtained. When using the protocol for mechanical isolation, the efficiency of isolating the islets of Langerhans in humans and experimental animals was 50%. The research proved that the isolated cells belonged to the islets of Langerhans. Additionally, their purity was confirmed. The survival of the isolated islets exceeded 90%. The enzyme immunoassay for insulin synthesis showed that the isolated islets of Langerhans remained functionally active. The sterility of the isolated islet cells was confirmed.

Conclusion. This proven isolation technology makes it possible to obtain pure, sterile, viable, and functionally active islet cells suitable for further autotransplantation.

1. Zureikat A.H., Nguyen T., Boone B.A., Wijkstrom M., Hogg M.E., Humar A., Zeh H. 3rd. Robotic total pancreatectomy with or without autologous islet cell trans plantation: replication of an open technique through a minimal access approach. Surg. Endosc. 2015; 29 (1): 176–183. https://doi.org/10.1007/s00464-014-3656-x

2. Sutherland D.E., Radosevich D.M., Bellin M.D., Hering B.J., Beilman G.J., Dunn T.B., Chinnakotla S., Vickers S.M., Bland B., Balamurugan A.N., Freeman M.L., Pruett T.L. Total pancreatectomy and islet autotransplantation for chronic pancreatitis. J. Am. Coll. Surg. 2012; 214 (4): 409–424; discussion 424–426. https://doi.org/10.1016/j.jamcollsurg.2011.12.040

3. Berger M., Beilman G.J., Dunn T., Sutherland D., Chinnakotla S. Total pancreatectomy and islet auto-transplantation for chronic painful pancreatitis: an overview. Int. J. Transplant. Res. Med. 2015; 1: 014. https://doi.org/10.23937/2572-4045.1510014

4. Zhang Y.J., Duan D.D., Yuan H. Efficacy and safety of islet autotransplantation after total pancreatectomy in chronic pancrea titis: a systematic review and meta-analysis including 17 studies. Clin. Res. Hepatol. Gastroenterol. 2020; 44 (4): 598–608. https://doi.org/10.1016/j.clinre.2019.08.004

5. Zagainov V.Е., Meleshina A.V., Korneva К.G., Vasenin S.А., Zagaynova E.V. Transplantation technologies for treatment of carbohydrate metabolism disorders. Russian Journal of Transplantology and Artificial Organs. 2020; 22 (1): 184–195. https://doi.org/10.15825/1995-1191-2020-1-184-195 (In Russian)

6. Bellin M.D., Balamurugan A.N., Pruett T.L., Sutherland D.E. No islets left behind: islet autotransplantation for surgeryinduced diabetes. Curr. Diab. Rep. 2012; 12 (5): 580–586. https://doi.org/10.1007/s11892-012-0296-1

7. Savari O., Golab K., Wang L.J., Schenck L., Grose R., Tibudan M., Ramachandran S., Chon W.J., Posner M.C., Millis J.M., Matthews J.B., Gelrud A., Witkowski P. Preservation of beta cell function after pancreatic islet autotransplantation: University of Chicago experience. Am. Surg. 2015; 81 (4): 421–427. PMID: 25831191

8. Fan C.J., Hirose K., Walsh C.M., Quartuccio M., Desai N.M., Singh V.K., Kalyani R.R., Warren D.S., Sun Z., Hanna M.N., Makary M.A. Laparoscopic total pancreatectomy with islet autotransplantation and intraoperative islet separation as a treatment for patients with chronic pancreatitis. JAMA Surg. 2017; 152 (6): 550–556. https://doi.org/10.1001/jamasurg.2016.5707

9. Aleotti F., Nano R., Piemonti L., Falconi M., Balzano G. Total pancreatectomy sequelae and quality of life: results of islet autotransplantation as a possible mitigation strategy. Updates Surg. 2021; 73 (4): 1237–1246. https://doi.org/10.1007/s13304-021-01129-w

10. Chaouch M.A., Leon P., Cassese G., Aguilhon C., Khayat S., Panaro F. Total pancreatectomy with intraportal islet autotransplantation for pancreatic malignancies: a literature overview. Expert. Opin. Biol. Ther. 2022; 22 (4): 491–497. https://doi.org/10.1080/14712598.2022.1990261

11. Jabłońska B., Mrowiec S. Total pancreatectomy with autologous islet cell transplantation – the current indications. J. Clin. Med. 2021; 10 (12): 2723. https://doi.org/10.3390/jcm10122723

12. Renaud F., Chetboun M., Thevenet J., Delalleau N., Gmyr V., Hubert T., Bonner C., Messager M., Leteurtre E., Mariette C., Kerr-Conte J., Piessen G., Pattou F. Safety of islet autotransplantation after pancreatectomy for adenocarcinoma. Transplantation. 2019; 103 (1): 177–181. https://doi.org/10.1097/TP.0000000000002419

13. Dugnani E., Pasquale V., Liberati D., Citro A., Cantarelli E., Pellegrini S., Marra P., Canu T., Balzano G., Scavini M., Esposito A., Doglioni C., Piemonti L. Modeling the iatrogenic pancreatic cancer risk after islet autotransplantation in mouse. Am. J. Transplant. 2017; 17 (10): 2720–2727. https://doi.org/10.1111/ajt.14360

14. Balzano G., Maffi P., Nano R., Zerbi A., Venturini M., Melzi R., Mercalli A., Magistretti P., Scavini M., Castoldi R., Carvello M., Braga M., Del Maschio A., Secchi A., Staudacher C., Piemonti L. Extending indications for islet autotransplantation in pancreatic surgery. Ann. Surg. 2013; 258 (2): 210–218. https://doi.org/10.1097/SLA.0b013e31829c790d

15. Yadav K.D., Huang X., Zhang Q., Zhang G., Bai X., Liang T. Auto-intestine transplantation for pancreatic tumors with mesenteric root involvement: a systematic review and survivalbased analysis. J. Pancreatol. 2021; 4 (4): 153–163. https://doi.org/10.1097/JP9.0000000000000081

16. Shapiro A.M., Pokrywczynska M., Ricordi C. Clinical pancreatic islet transplantation. Nat. Rev. Endocrinol. 2017; 13 (5): 268–277. https://doi.org/10.1038/nrendo.2016.178

17. Rheinheimer J., Bauer A.C., Silveiro S.P., Estivalet A.A., Bouças A.P., Rosa A.R., Souza B.M., Oliveira F.S., Cruz L.A., Brondani L.A., Azevedo M.J., Lemos N.E., Carlessi R., Assmann T.S., Gross J.L., Leitão C.B., Crispim D. Human pancreatic islet transplantation: an update and description of the establishment of a pancreatic islet isolation laboratory. Arch. Endocrinol. Metab. 2015; 59 (2): 161–170. https://doi.org/10.1590/2359-3997000000030

18. Ansite J., Balamurugan A.N., Barbaro B., Battle J., Brandhorst D., Cano J., Chen X., Deng S., Feddersen D., Friberg A., Gilmore T., Goldstein J.S., Holbrook E., Khan A., Kin T., Lei J., Linetsky E., Liu C., Luo X., McElvaney K., Min Z., Moreno J., O'Gorman D., Papas K.K., Putz G., Ricordi C., Szot G., Templeton T., Wang L., Wilhelm J.J., Willits J., Wilson T., Zhang X.: NIH CIT Consortium Chemistry Manufacturing Controls Monitoring Committee; NIH CIT Consortium. Purified Human Pancreatic Islets, CIT. Purification Density Gradients: A Standard Operating Procedure of the NIH Clinical Islet Transplantation Consortium. CellR4 Repair. Replace Regen. Reprogram. 2014; 2 (3): e991. PMID: 30613694; PMCID: PMC6319662

19. Corbin K.L., West H.L., Brodsky S., Whitticar N.B., Koch W.J., Nunemaker C.S. A practical guide to rodent islet isolation and assessment revisited. Biol. Proced. Online. 2021; 23 (1): 7. https://doi.org/10.1186/s12575-021-00143-x

20. Eich T., Ståhle M., Gustafsson B., Horneland R., Lempinen M., Lundgren T., Rafael E., Tufveson G., Zur-Mühlen B.V., Olerud J., Scholz H., Korsgren O. Calcium: a crucial potentiator for efficient enzyme digestion of the human pancreas. Cell Transplant. 2018; 27 (7): 1031–1038. https://doi.org/10.1177/0963689718779350

21. de Groot M., de Haan B.J., Keizer P.P., Schuurs T.A., van Schilfgaarde R., Leuvenink H.G. Rat islet isolation yield and function are donor strain dependent. Lab. Anim. 2004; 38 (2): 200–206. https://doi.org/10.1258/002367704322968885

22. Chinnakotla S., Beilman G.J., Dunn T.B., Bellin M.D., Freeman M.L., Radosevich D.M., Arain M., Amateau S.K., Mallery J.S., Schwarzenberg S.J., Clavel A., Wilhelm J., Robertson R.P., Berry L., Cook M., Hering B.J., Sutherland D.E., Pruett T.L. Factors predicting outcomes after a total pancreatectomy and islet autotransplantation lessons learned from over 500 cases. Ann. Surg. 2015; 262 (4): 610–622. https://doi.org/10.1097/SLA.0000000000001453