Human placental laminin: Role in neuronal differentiation, cell adhesion and proliferation

Abstract

Human placental extract has wound healing potential. Immuno-blots revealed presence of laminin in placental extract (70 ± 0.257 µg/ml; n=3). It was purified using immuno-affinity chromatography. SDS-PAGE and SE-HPLC indicated a188 kDa protein with some small peptides. Since placental laminin existed in its truncated form, its roles in cellular migration, differentiation and wound healing were verified. Induction of cellular migration and motility in rat fibroblasts were enhanced by placental laminin as observed from scratch wound assay. Promotion of neuronal differentiation of PC12 cells by placental laminin was observed by phase contrast microscopy. Confocal images showed presence of laminin on the cell surface and along the axonal processes. Significant interaction between integrin receptors and laminin responsible for cellular differentiation was demonstrated from co-localization experiments. Union between integrin receptor and its synthetic antagonist revealed retarded pattern of neurite outgrowth in laminin treated cells. Animal model studies revealed faster wound healing in the presence of placental laminin. Induction of re-epithelialization and angiogenesis in wound area by cellular proliferation and adhesion were observed. The cytokine levels showed an initial rise and gradual fall over the duration of wound healing on application of the fragmented laminin. Thus, roles of placental laminin in neuronal differentiation and wound healing were indicated.

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Abbreviations

ALP:

alkaline phosphatase

BCIP/NBT:

5-bromo-4-chloro-3-indolyl-phosphate/nitro blue tetrazolium

BSA:

bovine serum albumin

CRF:

corticotrophin releasing factor

EDC:

1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, HCl

Laminin:

the biomolecule as described in textbooks

IFN-ϒ:

interferon-ϒ

MES:

2-(N-morpholino) ethansulfonic acid

NADPH:

nicotinamide adenine dinucleotide phosphate

NBT:

nitro blue tetrazolium

NGF:

nerve growth factor

PBS:

phosphate buffered saline, pH 7.4

Placental laminin:

fragmented laminin from an aqueous extract from human placenta

pNPP:

para-nitrophenylphosphate

SGPT:

Serum glutamic pyruvic transaminase

SGOT:

serum glutamic oxaloacetic transaminase

STI:

soyabean trypsin inhibitor

TEMED:

N,N,N’,N’-tetramethylethane-1,2-diamine

TGF-β:

tumor growth factor

TNF-α:

tumor necrosis factor-α

Standard laminin:

full-length human placental laminin from Sigma

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Acknowledgements

CM and SS were Senior Research Fellows, CSIR, New Delhi, India. SD was supported by CSIR–Network Project miND-BSC 0115. The technical help rendered by Diptodeep Sirkar, Sounak Bhattacharya and Dipayan De is acknowledged for their contributions in confocal and super-resolution microscopy and quantification of confocal image measurements respectively.

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Correspondence to Debasish Bhattacharyya.

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Corresponding editor: Aurnab Ghose

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Mukherjee, C., Saleem, S., Das, S. et al. Human placental laminin: Role in neuronal differentiation, cell adhesion and proliferation. J Biosci 45, 93 (2020). https://doi.org/10.1007/s12038-020-00043-4

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Keywords

  • Cell adhesion
  • human placenta
  • laminin
  • neurite outgrowth
  • wound healing