DIFFERENTIATION OF BONE MARROW MESENCHYMAL STEM CELLS INTO INSULIN PRODUCING CELLS INDUCED BY RAT INJURED PANCREATIC TISSUE EXTRACT_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

FEIYun ^1,2 , XIEHongbin ² , WANGYunshuai ² , QIHui ¹ , DENGChunyan ² , RENLili ² ,  LIFurong ²

1. Department of Laboratory Medicine, the Second Clinic Medicine College(Shenzhen People's Hospital), Jinan University, Shenzhen Guangdong, 518020, P. R. China;
2. the Key Laboratory of Stem Cell and Cellular Therapy, the Second Clinic Medicine College(Shenzhen People's Hospital), Jinan University, Shenzhen Guangdong, 518020, P. R. China;

Corresponding author：

LIFurong, Email: frli62@163.com

Keywords：

Pancreas tissue engineering; Bone marrow mesenchymal stem cells; Insulin producing cells; Injured pancreatic tissue extract; Differentiation; Rat

DOI：

10.7507/1002-1892.20140139

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Objective To investigate the effects and mechanisms of differentiation of bone marrow mesenchymal stem cells (BMSCs) into insulin producing cells (IPCs) induced by injured pancreatic tissue extract of rat. Methods Eighty 6-week-old Sprague Dawley rats were selected. Forty rats underwent removal of 60% pancreas and the injured pancreas tissue was obtained after 48 hours to prepare the injured pancreatic tissue extract; and normal pancreatic tissue extract was prepared from the other 40 rats. The BMSCs were isolated from the tibia and femur of 4-week-old Sprague Dawley rats. BMSCs at passage 3 were co-cultured with rat injured pancreatic tissue extract as experimental group, with rat normal pancreatic tissue extract as normal control group, and with cell culture medium as blank control group for 14 days. The expressions of pancreas development related genes and proteins were detected, and cell morphological changes were observed. Then the C peptide positive cell rate was detected by flow cytometry analysis and insulin secretion levels were detected by glucose stimulation experiment at 14 days. Results Injured pancreatic tissue extract can induced BMSCs differentiating into IPCs. The pancreatic development related genes of pancreatic duodenal homeobox 1 (PDX-1), islet 1, Nkx6.1, glucose transporter type 2, proprotein convertase 2, neurogenin 3, and somatostatin were expressed sequentially in the differentiation process of experimental group; mature pancreatic proteins of PDX-1, insulin, C peptide, Nkx6.1 also were expressed. But there was no morphological changes and expression of pancreatic development related genes and proteins in normal control and blank control groups. The C peptide positive cell rate of experimental group (13.8%±1.8%) were significantly higher than those of normal control and blank control groups (1.6%±0.4%) (P<0.05). The insulin secretion of experimental group was significantly higher than that of normal control and blank control groups (P<0.05), but it was 1/40 and 1/47 of natural islet cells (P<0.05). Conclusion After pancreatic injury, injured pancreas would secrete transcription proteins related to development, differentiation, and repair of pancreas, which can promote the differentiation of BMSCs into IPCs.

Citation： FEIYun, XIEHongbin, WANGYunshuai, QIHui, DENGChunyan, RENLili, LIFurong. DIFFERENTIATION OF BONE MARROW MESENCHYMAL STEM CELLS INTO INSULIN PRODUCING CELLS INDUCED BY RAT INJURED PANCREATIC TISSUE EXTRACT. Chinese Journal of Reparative and Reconstructive Surgery, 2014, 28(5): 625-632. doi: 10.7507/1002-1892.20140139 Copy

1.	Tsai PJ, Wang HS, Lin CH, et al. Intraportal injection of insulin-producing cells generated from human bone marrow mesenchymal stem cells decreases blood glucose level in diabetic rats. Endocr Res, 2014, 39(1):26-33.
2.	Boroujeni Z, Aleyasin A. Human umblical cord derived mesenchymal stem cells can secret insulin in vitro and in vivo. Biotechnol Appl Biochem, 2013.[Epub ahead of print].
3.	Moshtagh PR, Emami SH, Sharifi AM. Differentiation of human adipose-derived mesenchymal stem cell into insulin-producing cells:an in vitro study. J Physiol Biochem, 2013, 69(3):451-458.
4.	Rahmati S, Alijani N, Kadivar M. In vitro generation of glucose-responsive insulin producing cells using lentiviral based pdx-1 gene transduction of mouse (C57BL/6) mesenchymal stem cells. Biochem Biophys Res Commun, 2013, 437(3):413-419.
5.	Tang DQ, Wang Q, Burkhardt BR, et al. In vitro generation of functional insulin-producing cells from human bone marrow-derived stem cells, but long-term culture running risk of malignant transformation. Am J Stem Cells, 2012, 1(2):114-127.
6.	Karaoz E, Okcu A, Ünal ZS, et al. Adipose tissue-derived mesenchymal stromal cells efficiently differentiate into insulin-producing cells in pancreatic islet microenvironment both in vitro and in vivo. Cytotherapy, 2013, 15(5):557-570.
7.	Peister A, Mellad JA, Larson BL, et al. Adult stem cells from bone marrow (MSCs) isolated from different strains of inbred mice vary in surface epitopes, rates of proliferation, and differentiation potential. Blood, 2004, 103(5):1662-1668.
8.	Hardikar AA, Karandikar MS, Bhonde RR. Effect of partial pancreatectomy on diabetic status in BALB/c mice. J Endocrinol, 1999, 162(2):189-195.
9.	Li FR, Qiu L, Yang XF, et al. The protective effects of ulinastatin and adenosine triphosphate on islet function during islet isolation and after islet transplantation. Pancreas, 2009, 38(2):227-230.
10.	Hardikar AA, Bhonde RR. Modulating experimental diabetes by treatment with cytosolic extract from the regenerating pancreas. Diabetes Res Clin Pract, 1999, 46(3):203-211.
11.	Kanitkar M, Bhonde R. Existence of islet regenerating factors within the pancreas. Rev Diabet Stud, 2004, 1(4):185-192.
12.	Choi KS, Shin JS, Lee JJ, et al. In vitro trans-differentiation of rat mesenchymal cells into insulin-producing cells by rat pancreatic extract. Biochem Biophys Res Commun, 2005, 330(4):1299-1305.
13.	Heinis M, Simon MT, Duvillié B. New insights into endocrine pancreatic development:the role of environmental factors. Horm Res Paediatr, 2010, 74(2):77-82.
14.	Chen W, Begum S, Opare-Addo L, et al. Promotion of beta-cell differentiation in pancreatic precursor cells by adult islet cells. Endocrinology, 2009, 150(2):570-579.
15.	Chang C, Niu D, Zhou H, et al. Mesenchymal stem cells contribute to insulin-producing cells upon microenvironmental manipulation in vitro. Transplant Proc, 2007, 39(10):3363-3368.
16.	Xie HB, Zhang H, Qi H, et al. Discovery of novel proteins from injured rat pancreatic extract using MALDI-TOF/MS-based proteomics. J Proteomics Bioinform, 2013, 6(8):158-163.
17.	Petropavlovskaia M, Rosenberg L. Indentification and characterization of small cell in the adult pancreas:Potential progenitor cells. Cell Tis Res, 2002, 310(1):51-58.
18.	Lumelsky N, Blondel O, Laeng P, et al. Differentiation of embryonic stem cells to insulin-secreting structures similar to pancreatic islets. Science, 2001, 292(5520):1389-1394.
19.	Börjesson A, Carlsson C. Altered proinsulin conversion in rat pancreatic islets exposed long-term to various glucose concentrations or interleukin-1beta. J Endocrinol, 2007, 192(2):381-387.

1. Tsai PJ, Wang HS, Lin CH, et al. Intraportal injection of insulin-producing cells generated from human bone marrow mesenchymal stem cells decreases blood glucose level in diabetic rats. Endocr Res, 2014, 39(1):26-33.
2. Boroujeni Z, Aleyasin A. Human umblical cord derived mesenchymal stem cells can secret insulin in vitro and in vivo. Biotechnol Appl Biochem, 2013.[Epub ahead of print].
3. Moshtagh PR, Emami SH, Sharifi AM. Differentiation of human adipose-derived mesenchymal stem cell into insulin-producing cells:an in vitro study. J Physiol Biochem, 2013, 69(3):451-458.
4. Rahmati S, Alijani N, Kadivar M. In vitro generation of glucose-responsive insulin producing cells using lentiviral based pdx-1 gene transduction of mouse (C57BL/6) mesenchymal stem cells. Biochem Biophys Res Commun, 2013, 437(3):413-419.
5. Tang DQ, Wang Q, Burkhardt BR, et al. In vitro generation of functional insulin-producing cells from human bone marrow-derived stem cells, but long-term culture running risk of malignant transformation. Am J Stem Cells, 2012, 1(2):114-127.
6. Karaoz E, Okcu A, Ünal ZS, et al. Adipose tissue-derived mesenchymal stromal cells efficiently differentiate into insulin-producing cells in pancreatic islet microenvironment both in vitro and in vivo. Cytotherapy, 2013, 15(5):557-570.
7. Peister A, Mellad JA, Larson BL, et al. Adult stem cells from bone marrow (MSCs) isolated from different strains of inbred mice vary in surface epitopes, rates of proliferation, and differentiation potential. Blood, 2004, 103(5):1662-1668.
8. Hardikar AA, Karandikar MS, Bhonde RR. Effect of partial pancreatectomy on diabetic status in BALB/c mice. J Endocrinol, 1999, 162(2):189-195.
9. Li FR, Qiu L, Yang XF, et al. The protective effects of ulinastatin and adenosine triphosphate on islet function during islet isolation and after islet transplantation. Pancreas, 2009, 38(2):227-230.
10. Hardikar AA, Bhonde RR. Modulating experimental diabetes by treatment with cytosolic extract from the regenerating pancreas. Diabetes Res Clin Pract, 1999, 46(3):203-211.
11. Kanitkar M, Bhonde R. Existence of islet regenerating factors within the pancreas. Rev Diabet Stud, 2004, 1(4):185-192.
12. Choi KS, Shin JS, Lee JJ, et al. In vitro trans-differentiation of rat mesenchymal cells into insulin-producing cells by rat pancreatic extract. Biochem Biophys Res Commun, 2005, 330(4):1299-1305.
13. Heinis M, Simon MT, Duvillié B. New insights into endocrine pancreatic development:the role of environmental factors. Horm Res Paediatr, 2010, 74(2):77-82.
14. Chen W, Begum S, Opare-Addo L, et al. Promotion of beta-cell differentiation in pancreatic precursor cells by adult islet cells. Endocrinology, 2009, 150(2):570-579.
15. Chang C, Niu D, Zhou H, et al. Mesenchymal stem cells contribute to insulin-producing cells upon microenvironmental manipulation in vitro. Transplant Proc, 2007, 39(10):3363-3368.
16. Xie HB, Zhang H, Qi H, et al. Discovery of novel proteins from injured rat pancreatic extract using MALDI-TOF/MS-based proteomics. J Proteomics Bioinform, 2013, 6(8):158-163.
17. Petropavlovskaia M, Rosenberg L. Indentification and characterization of small cell in the adult pancreas:Potential progenitor cells. Cell Tis Res, 2002, 310(1):51-58.
18. Lumelsky N, Blondel O, Laeng P, et al. Differentiation of embryonic stem cells to insulin-secreting structures similar to pancreatic islets. Science, 2001, 292(5520):1389-1394.
19. Börjesson A, Carlsson C. Altered proinsulin conversion in rat pancreatic islets exposed long-term to various glucose concentrations or interleukin-1beta. J Endocrinol, 2007, 192(2):381-387.

Chinese Journal of Reparative and Reconstructive Surgery

DIFFERENTIATION OF BONE MARROW MESENCHYMAL STEM CELLS INTO INSULIN PRODUCING CELLS INDUCED BY RAT INJURED PANCREATIC TISSUE EXTRACT

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