Int J Stem Cells.  2016 Nov;9(2):186-191. 10.15283/ijsc16044.

Regeneration of Bovine Mammary Gland in Immunodeficient Mice by Transplantation of Bovine Mammary Epithelial Cells Mixed with Matrigel

• Park HJ
• Lee WY
• Jeong HY
• Song H

• KMID: 2362512
• DOI: http://doi.org/10.15283/ijsc16044

Abstract

BACKGROUND AND OBJECTIVES
With the global demand for dairy protein for consumption growing annually, there has been increasing activity in the research field of dairy protein synthesis and production. From a manipulation perspective, it is more difficult to use live cattle for laboratory studies on the production of milk as well as of dairy protein such as casein, as compared with using laboratory animals like rodents. Therefore, we aimed to develop a mouse model of bovine mammary alveolar ducts for laboratory-scale studies. We studied the formation of the bovine mammary gland ductal structure by transplanting the MAC-T bovine alveolar cell line into mice.
METHODS AND RESULTS
MAC-T cells (1×10⁷) were suspended in Matrigel and injected into the dorsal tissue of 8-week-old male BALB/C nude mice. Histological analysis of tissue dissected from the MAC-T cell-transplanted mice after 6 weeks showed the typical morphology of the tubuloalveolar female gland, as well as glands made up of branching ducts that were surrounded by smooth muscle with small alveoli budding off the ducts. In addition, the epithelial markers CK14 and CK18 were expressed within the duct-like structure. Prolactin was detected in the duct interior in these CK14+ and CK18+ cells but not in the non-transplanted MAC-T cells.
CONCLUSIONS
These results showed that duct-like tissue had been successfully formed after 6 weeks of transplantation of the CK14+ and CK18+ MAC-T cells into mice dorsal tissue. This mouse model will be a useful tool for further research on the bovine mammary gland.

Keyword

Bovine mammary gland; MAC-T cell; BALB/C nude mice; CK14; CK18; Prolactin

MeSH Terms

Animals
Animals, Laboratory
Caseins
Cattle
Cell Line
Epithelial Cells*
Female
Humans
Male
Mammary Glands, Human*
Mice*
Mice, Nude
Milk
Muscle, Smooth
Prolactin
Regeneration*
Rodentia
Caseins
Prolactin

Figure

• 

  Fig. 1 CK14, CK18, and prolactin expression in bovine mammary gland. (A) Biopsied bovine mammary gland tissues stained with hematoxylin and eosin revealed a ductal structure. Immunostaining for CK14 (B), CK18 (C), and prolactin (D) showed strong positive expression of all three in the mammary gland duct. Bar=50 μm and 100 μm.

• 

  Fig. 2 CK14 and CK18 expression in MAC-T cells. (A) Image of MAC-T cell culture. Immunostaining for the basal cell marker CK14 (B) and the luminal cell marker CK18 (C) showed strongly positive staining of both in bovine MAC-T cells. Bar=50 μm

• 

  Fig. 3 CK14, CK18, and prolactin expression in MAC-T cell-transplanted mice tissue. (A) Images of MAC-T cell-transplanted mice tissue were taken after transplantation (n=5). (B) Hematoxylin and eosin-stained section of the MAC-T cell-transplanted tissue at 6 weeks after transplantation. The basal cell marker CK14 (C) and the luminal cell marker CK18 (D) were localized exclusively in the duct-like structures of the dorsal tissue. (E) These ducts were strongly positive for prolactin upon immunostaining. Bar=50 μm in all figures.

Reference

References

1. Ryman VE, Packiriswamy N, Sordillo LM. Role of endothelial cells in bovine mammary gland health and disease. Anim Health Res Rev. 2015; 16:135–149. DOI: 10.1017/S1466252315000158. PMID: 26303748.

2. Hovey RC, Trott JF, Vonderhaar BK. Establishing a framework for the functional mammary gland: from endocrinology to morphology. J Mammary Gland Biol Neoplasia. 2002; 7:17–38. DOI: 10.1023/A:1015766322258. PMID: 12160083.

3. Parmar H, Cunha GR. Epithelial-stromal interactions in the mouse and human mammary gland in vivo. Endocr Relat Cancer. 2004; 11:437–458. DOI: 10.1677/erc.1.00659. PMID: 15369447.

4. Huynh HT, Robitaille G, Turner JD. Establishment of bovine mammary epithelial cells (MAC-T): an in vitro model for bovine lactation. Exp Cell Res. 1991; 197:191–199. DOI: 10.1016/0014-4827(91)90422-Q. PMID: 1659986.

5. Fekete ÁA, Givens DI, Lovegrove JA. Can milk proteins be a useful tool in the management of cardiometabolic health? An updated review of human intervention trials. Proc Nutr Soc. 2016; 75:328–341. DOI: 10.1017/S0029665116000264. PMID: 27150497.

6. Hill DR, Newburg DS. Clinical applications of bioactive milk components. Nutr Rev. 2015; 73:463–476. DOI: 10.1093/nutrit/nuv009. PMID: 26011900. PMCID: 4560033.

7. Johnson TL, Tomanek L, Peterson DG. A proteomic analysis of the effect of growth hormone on mammary alveolar cell-T (MAC-T) cells in the presence of lactogenic hormones. Domest Anim Endocrinol. 2013; 44:26–35. DOI: 10.1016/j.domaniend.2012.08.001.

8. Lee HY, Heo YT, Lee SE, Hwang KC, Lee HG, Choi SH, Kim NH. Short communication: retinoic acid plus prolactin to synergistically increase specific casein gene expression in MAC-T cells. J Dairy Sci. 2013; 96:3835–3839. DOI: 10.3168/jds.2012-5945. PMID: 23587393.

9. McCoard SA, Hayashi AA, Sciascia Q, Rounce J, Sinclair B, McNabb WC, Roy NC. Mammary transcriptome analysis of lactating dairy cows following administration of bovine growth hormone. Animal. 2016; 1–10.

10. Buser AC, Obr AE, Kabotyanski EB, Grimm SL, Rosen JM, Edwards DP. Progesterone receptor directly inhibits β-casein gene transcription in mammary epithelial cells through promoting promoter and enhancer repressive chromatin modifications. Mol Endocrinol. 2011; 25:955–968. DOI: 10.1210/me.2011-0064. PMID: 21527503. PMCID: 3386529.

11. Martignani E, Eirew P, Accornero P, Eaves CJ, Baratta M. Human milk protein production in xenografts of genetically engineered bovine mammary epithelial stem cells. PLoS One. 2010; 5:e13372. DOI: 10.1371/journal.pone.0013372. PMID: 20976049. PMCID: 2957408.

12. Hair WM, Gubbay O, Jabbour HN, Lincoln GA. Prolactin receptor expression in human testis and accessory tissues: localization and function. Mol Hum Reprod. 2002; 8:606–611. DOI: 10.1093/molehr/8.7.606. PMID: 12087074.

13. Yang J, Wang R, Li H, Lv Q, Meng W, Yang X. Lentivirus mediated RNA interference of EMMPRIN (CD147) gene inhibits the proliferation, matrigel invasion and tumor formation of breast cancer cells. Cancer Biomark. 2016; 17:237–247. DOI: 10.3233/CBM-160636. PMID: 27434292.

14. Dores C, Dobrinski I. De novo morphogenesis of testis tissue: an improved bioassay to investigate the role of VEGF165 during testis formation. Reproduction. 2014; 148:109–117. DOI: 10.1530/REP-13-0303. PMID: 24803491. PMCID: 4130466.

15. Zhang H, Eisenried A, Zimmermann W, Shively JE. Role of CEACAM1 and CEACAM20 in an in vitro model of prostate morphogenesis. PLoS One. 2013; 8:e53359. DOI: 10.1371/journal.pone.0053359. PMID: 23358633. PMCID: 3554727.

16. Shultz LD, Schweitzer PA, Christianson SW, Gott B, Schweitzer IB, Tennent B, McKenna S, Mobraaten L, Rajan TV, Greiner DL, et al. Multiple defects in innate and adaptive immunologic function in NOD/LtSz-scid mice. J Immunol. 1995; 154:180–191. PMID: 7995938.

17. Líška J, Brtko J, Dubovický M, Macejová D, Kissová V, Polák Š, Ujházy E. Relationship between histology, development and tumorigenesis of mammary gland in female rat. Exp Anim. 2016; 65:1–9. DOI: 10.1538/expanim.15-0055.

18. Howlin J, McBryan J, Martin F. Pubertal mammary gland development: insights from mouse models. J Mammary Gland Biol Neoplasia. 2006; 11:283–297. DOI: 10.1007/s10911-006-9024-2. PMID: 17089203.

19. Hovey RC, Goldhar AS, Baffi J, Vonderhaar BK. Transcriptional regulation of vascular endothelial growth factor expression in epithelial and stromal cells during mouse mammary gland development. Mol Endocrinol. 2001; 15:819–831. DOI: 10.1210/mend.15.5.0635. PMID: 11328861.