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[Involvement of human amniotic fluid colony derived stem cells in regeneration of mouse injured muscle].

Abstract

OBJECTIVE

To study whether human amniotic fluid colony derived stem cells (hAFCSCs) are involved in regeneration of injured muscles in mice and to investigate the method and feasibility of hAFCSCs-based cytotherapy in the treatment of injured muscles.

METHODS

s Human second-trimester amniotic fluid was collected through ultrasound-guided amniocentesis, hAFCSCs were isolated from second-trimester amniotic fluid and cultured, and the cells at 6th-8th passages were spared. The mRNA was extracted to identify the stem cell related genes by RT-PCR. The muscular injury model of bilateral tibialis anterior muscle was established by cardiotoxin and X-ray irradiation in 16 Nod/Scid mice (aged 6-8 weeks, and weighing 20-24 g). The hAFCSCs (3.3 x 1(0)7/mL, 3 microIL) were injected into the right injured tibialis anterior muscles as the experimental group, while the same volume of complete medium (lphaa-MEM containing 15%FBS, 18%Chang B, 2%Chang C, 1% penicillin-streptomycin, and 1% L-glutamine) was injected into the left injured tibialis anterior muscles as the control group. At 2 and 4 weeks after cell transplantation, the immunofluorescence staining of tibialis anterior muscles was performed to detect hepatocyte growth factor receptor (c-Met), myogenic regulatory factor (Myf-5), Laminin, Desmin, and human specific nuclear mitotic apparatus protein (NuMa).

RESULTS

s The clone formation was observed at 5-7 days of primary hAFCSCs culture; after 8-10 days, the clones with homogeneous morphology were selected for subculture. Adequate stem cells were available after 6th-8th subculture. RT-PCR analysis showed that hAFCSCs expressed mRNA of the stem cell related genes. The immunofluorescence double-staining showed that NuMa expressed in tibialis anterior muscles of the experimental group and no myogenic phenotype expressed at 2 weeks after cell transplantation, and that single cell co-expressed NuMa and c-Met or Myf-5 at 4 weeks after cell transplantation. In some myofibers, NuMa and Laminin or Desmin were also co-expressed. No NuMa positive hAFCSCs were detected in the control group at 2 and 4 weeks after cell transplantation.

CONCLUSION

n hAFCSCs can participate in the regeneration of injured mouse muscle.

Authors+Show Affiliations

Central Laboratory, Shanghai Children's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, 200040, P.R.China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

English Abstract
Journal Article
Research Support, Non-U.S. Gov't

Language

chi

PubMed ID

21818952

Citation

Ma, Xiaorong, et al. "[Involvement of Human Amniotic Fluid Colony Derived Stem Cells in Regeneration of Mouse Injured Muscle]." Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi = Zhongguo Xiufu Chongjian Waike Zazhi = Chinese Journal of Reparative and Reconstructive Surgery, vol. 25, no. 7, 2011, pp. 842-7.
Ma X, Zhang S, Shang Y, et al. [Involvement of human amniotic fluid colony derived stem cells in regeneration of mouse injured muscle]. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2011;25(7):842-7.
Ma, X., Zhang, S., Shang, Y., Gao, T., Wang, X., Chen, F., & Zhou, J. (2011). [Involvement of human amniotic fluid colony derived stem cells in regeneration of mouse injured muscle]. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi = Zhongguo Xiufu Chongjian Waike Zazhi = Chinese Journal of Reparative and Reconstructive Surgery, 25(7), 842-7.
Ma X, et al. [Involvement of Human Amniotic Fluid Colony Derived Stem Cells in Regeneration of Mouse Injured Muscle]. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2011;25(7):842-7. PubMed PMID: 21818952.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Involvement of human amniotic fluid colony derived stem cells in regeneration of mouse injured muscle]. AU - Ma,Xiaorong, AU - Zhang,Shengli, AU - Shang,Yafeng, AU - Gao,Tongbin, AU - Wang,Xue, AU - Chen,Fang, AU - Zhou,Junmei, PY - 2011/8/9/entrez PY - 2011/8/9/pubmed PY - 2012/4/20/medline SP - 842 EP - 7 JF - Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = Chinese journal of reparative and reconstructive surgery JO - Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi VL - 25 IS - 7 N2 - OBJECTIVE: To study whether human amniotic fluid colony derived stem cells (hAFCSCs) are involved in regeneration of injured muscles in mice and to investigate the method and feasibility of hAFCSCs-based cytotherapy in the treatment of injured muscles. METHODS: s Human second-trimester amniotic fluid was collected through ultrasound-guided amniocentesis, hAFCSCs were isolated from second-trimester amniotic fluid and cultured, and the cells at 6th-8th passages were spared. The mRNA was extracted to identify the stem cell related genes by RT-PCR. The muscular injury model of bilateral tibialis anterior muscle was established by cardiotoxin and X-ray irradiation in 16 Nod/Scid mice (aged 6-8 weeks, and weighing 20-24 g). The hAFCSCs (3.3 x 1(0)7/mL, 3 microIL) were injected into the right injured tibialis anterior muscles as the experimental group, while the same volume of complete medium (lphaa-MEM containing 15%FBS, 18%Chang B, 2%Chang C, 1% penicillin-streptomycin, and 1% L-glutamine) was injected into the left injured tibialis anterior muscles as the control group. At 2 and 4 weeks after cell transplantation, the immunofluorescence staining of tibialis anterior muscles was performed to detect hepatocyte growth factor receptor (c-Met), myogenic regulatory factor (Myf-5), Laminin, Desmin, and human specific nuclear mitotic apparatus protein (NuMa). RESULTS: s The clone formation was observed at 5-7 days of primary hAFCSCs culture; after 8-10 days, the clones with homogeneous morphology were selected for subculture. Adequate stem cells were available after 6th-8th subculture. RT-PCR analysis showed that hAFCSCs expressed mRNA of the stem cell related genes. The immunofluorescence double-staining showed that NuMa expressed in tibialis anterior muscles of the experimental group and no myogenic phenotype expressed at 2 weeks after cell transplantation, and that single cell co-expressed NuMa and c-Met or Myf-5 at 4 weeks after cell transplantation. In some myofibers, NuMa and Laminin or Desmin were also co-expressed. No NuMa positive hAFCSCs were detected in the control group at 2 and 4 weeks after cell transplantation. CONCLUSION: n hAFCSCs can participate in the regeneration of injured mouse muscle. SN - 1002-1892 UR - https://www.unboundmedicine.com/medline/citation/21818952/[Involvement_of_human_amniotic_fluid_colony_derived_stem_cells_in_regeneration_of_mouse_injured_muscle]_ L2 - https://medlineplus.gov/stemcells.html DB - PRIME DP - Unbound Medicine ER -