Hostname: page-component-586b7cd67f-g8jcs Total loading time: 0 Render date: 2024-11-27T00:53:09.008Z Has data issue: false hasContentIssue false
  • English
  • Français

Modulation of the viability of immature cardiac myocytes by cardiac fibroblasts after hypothermic preservation—its values as a technique for evaluation of storage solutions

Published online by Cambridge University Press:  19 August 2008

Hiroyuki Orita ,
Manabu Fukasawa ,
Hideaki Uchino ,
Kana Fukui ,
Minoru Kohi  and
Masahiko Washio
Hiroyuki Orita*
Affiliation:
From the The Second Department of Surgery, Yamagata University School of Medicine, Yamagata City
Manabu Fukasawa
Affiliation:
From the The Second Department of Surgery, Yamagata University School of Medicine, Yamagata City
Hideaki Uchino
Affiliation:
From the The Second Department of Surgery, Yamagata University School of Medicine, Yamagata City
Kana Fukui
Affiliation:
From the The Second Department of Surgery, Yamagata University School of Medicine, Yamagata City
Minoru Kohi
Affiliation:
From the The Second Department of Surgery, Yamagata University School of Medicine, Yamagata City
Masahiko Washio
Affiliation:
From the The Second Department of Surgery, Yamagata University School of Medicine, Yamagata City
*
Dr. Hiroyuki Orita, The Second Department of Surgery, Yamagata University School of Medicine, lida-nishi, Yamagata City, 990–23, Japan. Tel. 81-236-33-1235; Fax. 81-236-25-9122.
Get access Rights & Permissions [Opens in a new window]

Abstract

We evaluated the modulation of the viability of immature cardiac myocytes by cardiac fibroblasts after hypothermic preservation using three types of storage solutions—saline, University of Wisconsin solution, and MCDB 107 medium. Cardiac myocytes and fibroblasts were isolated from neonatal rat ventricles, and cultures of myocytes only or co-cultures with fibroblasts (myocyte: fibroblast 2:1) were established. On the fourth day of culture, the cultures were incubated at 4 °C for 6, 12, 18 and 24 hours in the different storage solutions. Enzymes were measured in the storage solutions immediately before and after hypothermic incubation. The cultures were then incubated for an additional 24 hours at 37 °C to evaluate the recovery of the myocyte beating rate. The myocyte beating rate in the co-culture groups showed significantly higher recovery ratios than the corresponding groups in which only myocytes were cultured. Complete recovery was observed in the group co-cultured in MCDB medium 24 hours after hypothermic incubation (83.4% of control—beating rate prior to hypothermic incubation) compared to the other co-cultured groups (15.4, 0%, respectively). Release of enzymes in the co-cultures was significantly suppressed compared to the cultured myocytes, and the greatest suppression was found after 24 hours of incubation in MCDB medium (CPK: 36.6 mIU/flask, LDH: 281.2 mIU/flask) compared to the other two co-cultured groups (CPK: 181.1, 281.1; LDH: 501.7, 773.2). Cardiac fibroblasts diminished myocytic injury after hypothermic preservation using various storage solutions, in which MCDB 107 medium showed the best overall protective effect. Thus, cardiac fibroblasts may play an important role in controlling myocytic viability under various hypothermic conditions.

Keywords

Cardiac myocytecardiac fibroblasthypothermic injuryMCDB 107 mediumUniversity of Wisconsin solution
Type
Original Articles
Information
Cardiology in the Young , Volume 5 , Issue 2 , April 1995 , pp. 110 - 117
Copyright
Copyright © Cambridge University Press 1995

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

1.Burt, JM, Copeland, JG.Myocardial function after preservation for 24 hours. J Thorac Cardiovasc Surg 1986; 92: 238–246.CrossRefGoogle ScholarPubMed
2.Takahashi, A, Braimbridge, MV, Hearse, DJ, Chambers, DJ.Long-term preservation of the mammalian myocardium. Effect of storage medium and temperature on the vulnerability to tissue injury. J Thorac Cardiovasc Surg 1991; 102: 235–244.CrossRefGoogle ScholarPubMed
3.Wicomb, W, Cooper, DKC, Hassoulas, J, Rose, AG, Barnard, CN.Orthotopic transplantation of the baboon heart after 20 to 24 hours preservation by continuous hypothermic perfusion with an oxygenated hyperosmolar solution. J Thorac Cardiovasc Surg 1982; 83: 133–140.CrossRefGoogle ScholarPubMed
4.Warner, M, Guerraty, A, Alivizatos, P, Choi, SC, Hudson, B, Lower, RR, Hess, ML.Assessment of myocardial subcellular function after 24 hours of in vitro preservation and transplantation. J Thorac Cardiovasc Surg 1982; 83: 290–297.Google ScholarPubMed
5.Jarmakani, JM, Nakazawa, M, Nagatomo, T, Langer, GA.Effect of hypoxia on mechanical function in the neonatal mammalian heart. Am J Physiol 1978; 235: H469–H474.Google ScholarPubMed
6.Bove, EL, Gallagher, KP, Drake, DH, Lynch, MJ, Fox, M, Forder, J, Boling, SF, Shlafer, M.The effect of hypothermic ischemia on recovery of left ventricular function and preload reserve in the neonatal heart. J Thorac Cardiovasc Surg 1988; 95: 814–818.CrossRefGoogle ScholarPubMed
7.Grice, WN, Konishi, T, Apstein, CS.Resistance of neonatal myocardium to injury during normothermic and hypothermic ischemic arrest and reperfusion. Circulation 1987; 76(Suppl. Vyes): V150–V155.Google ScholarPubMed
8.Orita, H, Fukasawa, M, Hirooka, S, Uchino, H, Fukui, K, Kohi, M, Washio, M.Cardiac myocyte functional and biochemical changes after hypothermic preservation in vitro: Protective effects of storage solutions. J Thorac Cardiovasc Surg 1994; 107: 226–232.CrossRefGoogle ScholarPubMed
9.Klein, I, Daood, M.Regulation of the growth of nonmuscle heart cells in culture. In Vitro Cell Dcv Biol 1985; 21: 693–696.CrossRefGoogle ScholarPubMed
10.Suzuki, T, Ohta, M, Hoshi, H.Serum-free, chemically defined medium to evaluate the direct effects of growth factors and inhibitors on proliferation and function of neonatal rat cardiac muscle cells in culture. In Vitro Cell Dev Biol 1989; 25: 601–606.CrossRefGoogle ScholarPubMed
11.Orita, H, Fukasawa, M, Hirooka, S, Fukui, K, Kohi, M, Washio, M.A cardiac myocyte culture system as an in vitro experimental model for the evaluation of hypothermic preservation. Surgery Today 1993; 23: 439–443.CrossRefGoogle Scholar
12.Orita, H, Fukasawa, M, Hirooka, S, Fukui, K, Kohi, M, Washio, M.Protection of cardiac myocytes from hypothermic injury by cardiac fibroblasts isolated from neonatal rat ventricle. J Surg Res 1993; 55: 654–658.CrossRefGoogle ScholarPubMed
13.Orita, H, Fukasawa, M, Hirooka, S, Uchino, H, Fukui, K, Kohi, M, Washio, M.Modulation of cardiac myocyte beating rate and hypertrophy by cardiac fibroblasts isolated from neonatal rat ventricle. Jpn Circ J 1993; 57: 912–920.CrossRefGoogle ScholarPubMed
14.Blondel, B, Roijen, I, Cheneval, JP.Heart cells in culture: a simple method for increasing the population of myoblasts. Experiential 1971; 27: 356–358.CrossRefGoogle Scholar
15.Polinger, IS.Separation of cell types in embryonic heart cell culture. Exp Cell Res 1970; 63: 78–82.CrossRefGoogle Scholar
16.Yagev, S, Heller, M, Pinson, A.Change in cytoplasmic and lysosomal enzyme activities in cultured rat heart cells. In Vitro Cell Dev Biol 1984; 20: 893–898.Google ScholarPubMed
17.Siegel, S, Castellan, NJ Jr. The Kruskal-Wallis one-way analysis of variance by ranks. In: Anker, JD (ed). Nonparametric Statistics for the Behavioral Sciences. McGraw-Hill Inc., New York, 1988, pp 206–216.Google Scholar
18.Siegel, S, Castellan, NJ Jr. The Wilcoxon-Mann-WhitneyTest. In: Anker, JD (ed). Nonparametric Statistics for the Behavioral Sciences. McGraw-Hill Inc., New York, 1988, pp 206–216.Google Scholar
19.Hendry, PJ, Labow, RS, Barry, TA, Keon, WJ.An assessment of crystalloid solutions for donor heart preservation. J Thorac Cardiovasc Surg 1991; 101: 833–838.CrossRefGoogle ScholarPubMed
20.Fremes, SE, Li, RK, Weisel, RD, Mickle, DAG, Tumiati, LC.Prolonged hypothermic cardiac storage with University of Wisconsin solution. An assessment with human cell culture. J Thorac Cardiovasc Surg 1991; 102: 666–672.CrossRefGoogle Scholar
21.Orita, H, Fukasawa, M, Hirooka, S, Uchino, H, Fukui, K, Kohi, M, Washio, M.In vitro evaluation of phosphate, bicarbonate and hepes buffered storage solutions on hypothermic injury to immature myocytes. Cardiovasc Drugs Ther [In press]Google Scholar
22.Wahlberg, JA, Love, R, Landegaard, L, Belzer, FO.72 hour preservation of the canine pancreas. Transplantation 1987; 43: 5–8.CrossRefGoogle ScholarPubMed
23.Schmid, T, Landry, G, Fields, BL, Belzer, FO, Haworth, RA, Southard, JH.The use of myocytes as a model for developing successful heart preservation solutions. Transplantation 1991; 52: 20–26.CrossRefGoogle Scholar
24.Page, E, Manjuath, CK. Communicating junctions between cardiac cells. In: Fozzard, HA, Haber, E, Jennings, RB, Katz, AM, Morgan, HE (eds). The Heart and Cardiovascular System. Raven Press, New York, 1986, pp 537–600.Google Scholar
25.Severs, NJ, Shovel, KS, Slade, AM, Powell, T, Twist, VW, Green, CR.Fate of gap junctions in isolated adult mammalian cardiomyocytes. Circ Res 1989; 65: 22–42.Google ScholarPubMed
26.Luke, RA, Beyer, EC, Hoyt, RH, Saffitz, JE.Quantitative analysis of intracellular connections by immunohistochemistry of the cardiac gap junction protein connexin-43. Circ Res 1989; 65: 1450–1457.CrossRefGoogle Scholar
27.Brog, TK.Specific attachment of collagen to cardiac myocytes. Devl Biol 1983; 97: 417–423.CrossRefGoogle Scholar
28.Eghbali, M, Czaja, MJ, Zeydel, M, Weiner, FR, Zern, MA, Seifter, S, Blumenfeld, OO.Collagen chains mRNAs in isolated heart cells from young and adult rats. J Mol Cell Cardiol 1988; 20: 267–276.CrossRefGoogle ScholarPubMed
29.Blumenfeld, OO, Seifter, S. Biochemistry of connective tissue with special emphasis on the heart. In: Robinson, TF, Kinne, RKH (eds). Cardiac Myocyte-Connective Tissue Interactions in Health and Disease. Karger, Basel, 1990, pp 5–36.Google Scholar
30.Brog, TK, Terracio, L. Interaction of the extracellular matrix with cardiac myocytes during development and disease. In: Robinson, TF, Kinne, RKH (eds). Cardiac Myocyte-Connective Tissue Interactions in Health and Disease. Kargerz, Basel, 1990, pp 113–129.Google Scholar