Isolation, Characterization and Osteogenic Potential of Mouse Digit Tip Blastema Cells in Comparison with Bone Marrow-Derived Mesenchymal Stem Cells In Vitro

Overview

Journal Cell J

Specialty Cell Biology

Date 2017 Nov 7

PMID 29105393

Citations 4

Authors

Leila Taghiyar

Samaneh Hosseini

Mahdi Hesaraki

Forough Azam Sayahpour

Nasser Aghdami

Mohamadreza Baghaban Eslaminejad

Affiliations

Soon will be listed here.

Abstract

Objectives: Limb regeneration mediated by blastema cells (BlCs) in mammals is limited to the digit tips of neonates. Due to the lack of access to BlCs in adults and the difficulty in isolating and expanding BlCs from neonates, the use of a cellular population with similar features of BlCs would be a valuable strategy to direct a non-regenerative wound towards regeneration. In this study, we have initially isolated and cultured BlCs, and explored their characteristics in vitro. Next, we compared the capability of bone marrow-derived mesenchymal stem cells (BM-MSCs) as an alternative accessible cell source to BlCs for regeneration of appendages.

Materials And Methods: In this experimental study, BM-MSCs were isolated from BM and we obtained BlCs from the neonatal regenerating digit tip of C57B/6 mice. The cells were characterized for expressions of cell surface markers by flow cytometry. Quantitative-reverse transcription polymerase chain reaction (qRT-PCR) and lineage-specific staining were used to assess their ability to differentiate into skeletal cell lineages. The colony forming ability, proliferation, alkaline phosphatase (ALP) activity, calcium content, and osteogenic gene expression were evaluated in both BMMSCs and BlCs cultures at days 7, 14, and 21.

Results: qRT-PCR analysis revealed that the cells from both sources readily differentiated into mesodermal lineages. There was significantly higher colony forming ability in BM-MSCs compared to BlCs (P<0.05). Alizarin red staining (ARS), calcium, and the ALP assay showed the same degree of mineral deposition in both BlCs and BM-MSCs. Gene expression levels of osteblastic markers indicated similar bone differentiation capacity for both BlCs and BM-MSCs at all time-points.

Conclusions: Characteristics of BlCs in vitro appear to be similar to BM-MSCs. Therefore, they could be considered as a substitute for BlCs for a regenerative approach with potential use in future clinical settings for regenerating human appendages.

Citing Articles

Donor age effects on in vitro chondrogenic and osteogenic differentiation performance of equine bone marrow- and adipose tissue-derived mesenchymal stromal cells.

Bagge J, Berg L, Janes J, MacLeod J BMC Vet Res. 2022; 18(1):388.

PMID: 36329434 PMC: 9632053. DOI: 10.1186/s12917-022-03475-2.

Automated digital image quantification of histological staining for the analysis of the trilineage differentiation potential of mesenchymal stem cells.

Eggerschwiler B, Canepa D, Pape H, Casanova E, Cinelli P Stem Cell Res Ther. 2019; 10(1):69.

PMID: 30808403 PMC: 6390603. DOI: 10.1186/s13287-019-1170-8.

Neonatal mouse intervertebral discs heal with restored function following herniation injury.

Torre O, Das R, Berenblum R, Huang A, Iatridis J FASEB J. 2018; 32(9):4753-4762.

PMID: 29570392 PMC: 6103171. DOI: 10.1096/fj.201701492R.

Msh homeobox 1 ()- and -overexpressing bone marrow-derived mesenchymal stem cells resemble blastema cells and enhance regeneration in mice.

Taghiyar L, Hesaraki M, Sayahpour F, Satarian L, Hosseini S, Aghdami N J Biol Chem. 2017; 292(25):10520-10533.

PMID: 28461333 PMC: 5481560. DOI: 10.1074/jbc.M116.774265.

References

Jeon O, Rhie J, Kwon I, Kim J, Kim B, Lee S . In vivo bone formation following transplantation of human adipose-derived stromal cells that are not differentiated osteogenically. Tissue Eng Part A. 2008; 14(8):1285-94. DOI: 10.1089/ten.tea.2007.0253. View

Takeo M, Chou W, Sun Q, Lee W, Rabbani P, Loomis C . Wnt activation in nail epithelium couples nail growth to digit regeneration. Nature. 2013; 499(7457):228-32. PMC: 3936678. DOI: 10.1038/nature12214. View

Bryant S, Endo T, Gardiner D . Vertebrate limb regeneration and the origin of limb stem cells. Int J Dev Biol. 2002; 46(7):887-96. View

Muneoka K, Sassoon D . Molecular aspects of regeneration in developing vertebrate limbs. Dev Biol. 1992; 152(1):37-49. DOI: 10.1016/0012-1606(92)90154-9. View

Yu L, Han M, Yan M, Lee E, Lee J, Muneoka K . BMP signaling induces digit regeneration in neonatal mice. Development. 2010; 137(4):551-9. PMC: 2827613. DOI: 10.1242/dev.042424. View

Bajek A, Gurtowska N, Olkowska J, Kazmierski L, Maj M, Drewa T . Adipose-Derived Stem Cells as a Tool in Cell-Based Therapies. Arch Immunol Ther Exp (Warsz). 2016; 64(6):443-454. PMC: 5085986. DOI: 10.1007/s00005-016-0394-x. View

Fernando W, Leininger E, Simkin J, Li N, Malcom C, Sathyamoorthi S . Wound healing and blastema formation in regenerating digit tips of adult mice. Dev Biol. 2010; 350(2):301-10. PMC: 3031655. DOI: 10.1016/j.ydbio.2010.11.035. View

Tanaka E . Cell differentiation and cell fate during urodele tail and limb regeneration. Curr Opin Genet Dev. 2003; 13(5):497-501. DOI: 10.1016/j.gde.2003.08.003. View

Phan A, Lee J, Oei M, Flath C, Hwe C, Mariano R . Positional information in axolotl and mouse limb extracellular matrix is mediated via heparan sulfate and fibroblast growth factor during limb regeneration in the axolotl (Ambystoma mexicanum). Regeneration (Oxf). 2016; 2(4):182-201. PMC: 4857728. DOI: 10.1002/reg2.40. View

10.

Hattori H, Sato M, Masuoka K, Ishihara M, Kikuchi T, Matsui T . Osteogenic potential of human adipose tissue-derived stromal cells as an alternative stem cell source. Cells Tissues Organs. 2004; 178(1):2-12. DOI: 10.1159/000081088. View

11.

Kawasumi A, Sagawa N, Hayashi S, Yokoyama H, Tamura K . Wound healing in mammals and amphibians: toward limb regeneration in mammals. Curr Top Microbiol Immunol. 2012; 367:33-49. DOI: 10.1007/82_2012_305. View

12.

Cai T, Zhu W, Chen X, Zhou S, Jia L, Sun Y . Fibroblast growth factor 2 induces mesenchymal stem cells to differentiate into tenocytes through the MAPK pathway. Mol Med Rep. 2013; 8(5):1323-8. DOI: 10.3892/mmr.2013.1668. View

13.

Penfornis P, Pochampally R . Isolation and expansion of mesenchymal stem cells/multipotential stromal cells from human bone marrow. Methods Mol Biol. 2011; 698:11-21. DOI: 10.1007/978-1-60761-999-4_2. View

14.

Muller I, Kordowich S, Holzwarth C, Isensee G, Lang P, Neunhoeffer F . Application of multipotent mesenchymal stromal cells in pediatric patients following allogeneic stem cell transplantation. Blood Cells Mol Dis. 2007; 40(1):25-32. DOI: 10.1016/j.bcmd.2007.06.021. View

15.

Rostovskaya M, Anastassiadis K . Differential expression of surface markers in mouse bone marrow mesenchymal stromal cell subpopulations with distinct lineage commitment. PLoS One. 2012; 7(12):e51221. PMC: 3517475. DOI: 10.1371/journal.pone.0051221. View

16.

Duque G . Bone and fat connection in aging bone. Curr Opin Rheumatol. 2008; 20(4):429-34. DOI: 10.1097/BOR.0b013e3283025e9c. View

17.

Franceschi R, Xiao G . Regulation of the osteoblast-specific transcription factor, Runx2: responsiveness to multiple signal transduction pathways. J Cell Biochem. 2003; 88(3):446-54. DOI: 10.1002/jcb.10369. View

18.

Soleimani M, Nadri S . A protocol for isolation and culture of mesenchymal stem cells from mouse bone marrow. Nat Protoc. 2009; 4(1):102-6. DOI: 10.1038/nprot.2008.221. View

19.

Lehoczky J, Robert B, Tabin C . Mouse digit tip regeneration is mediated by fate-restricted progenitor cells. Proc Natl Acad Sci U S A. 2011; 108(51):20609-14. PMC: 3251149. DOI: 10.1073/pnas.1118017108. View

20.

Sousa S, Afonso N, Bensimon-Brito A, Fonseca M, Simoes M, Leon J . Differentiated skeletal cells contribute to blastema formation during zebrafish fin regeneration. Development. 2011; 138(18):3897-905. DOI: 10.1242/dev.064717. View